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Photo by Jayne Kamin-Oncea/USA Today Sports
By MARK EDWARD HORNISH
CARSON, Calif. — If the LA Galaxy set out to send a message to the rest of Major League Soccer, it went something like this: "Be afraid. Be very afraid."
The Galaxy harshly welcomed D.C. United on Wednesday night, crushing the Eastern Conference leaders, 4-1, before 16,044 at the Stub Hub Center. The Galaxy spread the love, with goals coming from Alan Gordon, Omar Gonzalez, Baggio Husidic, and Landon Donovan.
Donovan once again figured prominently in the match, earning assists on both the Gordan and Gonzalez goals, drawing him within eight of Steve Ralston's career MLS record of 127. Donovan completed his evening — and stamped out any lingering DC United hopes — by converting a penalty kick in the 75th. He was subbed out in the 86th minute to a standing ovation.
"I'm enjoying it — I realize very clearly there's only a very few more chances to do this," Donovan said after the match. "The decision to retire, and playing this way, are intricately linked. That's given me freedom to relax and enjoy it, and it's taken all the pressure off. That's when I play my best."
"Landon's obviously committed to completing his career in the right manner," added Galaxy head coach Bruce Arena. "His leadership and quality tonight was terrific. What more can you say. He's been a great one. His attitude has been terrific over the last two or three weeks since his announcement. We're happy for him. He's been a big part of our success in over the last few months."
While the Husidic goal may not get goal-of-the-week attention, it was nonetheless extraordinary. The Galaxy strung together 26 consecutive passes before Husidic tapped home an AJ De La Garza cross to close out the first half.
"We've been scoring early in the game, which opens up the field for us," Husidic said after the match. "Another great team goal. A lot of connected passes. Very fluid soccer tonight. I don't know how many connected passes… twenty six? I think that's pretty impressive."
D.C. United showed some life early in the second half, stringing together a rapid sequence of shots, culminating in a goal created by, if not credited to, a hustling Eddie Johnson. Johnson beat beleaguered Galaxy center back Leonardo to the near post, allowing an Alex Caskey cross to hop over Galaxy goalkeeper Jaime Penedo. The goal was ruled an own goal by Leonardo.
Recent addition Alan Gordon started up top in place of Robbie Keane, who enjoyed the match in street clothes from David Beckham's luxury box. Gordon outdid Donovan's 4th minute goal in the previous match by scoring in the 2nd minute off a deflected Donovan shot. The early lead put United on the back foot for the remainder of the half.
Omar Gonzalez figured in the offense as well, punching home a Donovan cross in the 25th minute to double the Galaxy lead.
"It's a lot of fun playing right now. I'm trying to get forward as much as possible," Gonzalez said. "It was a very important win for us, gets us that much closer to the top of the table."
With the win and the three points, the Galaxy now vault into second place in the Western Conference, two behind the Seattle Sounders, and still with one game in-hand. In addition, the Galaxy now draw into a tie for second in the Supporter's Shield race.
When asked if the Galaxy are targeting the Supporter's Shield, Donovan responded without hesitation: "Yeah. We are. We wanna make a push for it. It's a little dangerous, when you have games in hand all season, because you add up the points in your head, without the points being on the board. But now we've won these games.
"We've leap-frogged Dallas and Salt Lake. We play Seattle twice more, and Dallas twice more. So, there's every chance that we can get there. And like we've been doing, we'll take it one game at a time."
The Galaxy remain at home, and will face Chivas USA on Sunday evening in the final edition ever of the Superclassico.
Despite the loss, DC United remain in first place in the Eastern Conference, while in the midst of their toughest stretch of the season. They return home and will face New York Red Bulls on Sunday.
Watch the match highlights below:
Klinsmann's balls must really hurt.
I agree… at the time, I was Ok with leaving him off the WC Squad… but looking back, we could have used him in Brazil.
And I love how he's been beast mode since being cut…I hope he gets the All Time Assist Record too..
It was never okay to leave him off the WC squad and pretty much everyone who knows anything about futbol outside of the USA thought it was wrong.
Diego's Maradoughnuts
I could care less what they think. Most of these people who "know futbol outside the US" couldn't name 10 current American players and wouldn't be caught dead watching an MLS game, assuming they could find one. They all picked us dead last in the group and they'll do it again. Who cares?
I didn't love the decision either but I don't need some Eurosnot telling us what happens in our house. They don't know squat about American soccer because none of them follow it. And most of them don't bathe regularly.
Quit Whining About Soccer in the US
DM with the post of the week, the month, the year.
insecure Americans need to quit worrying about what others think.
We are big boys now, we can have our own opinions and we can know they are better opinions.
If we are big boys and can have our own opinions then why do we tolerate a coach who continues to insult our intelligence, our soccer history, culture, and players (not just Donovan – Dempsey, Bradley, all the non Germans). Why do we let Klinsmann constantly tell us everything that is wrong with us while he basically tries to just emulate what Germany does rather than try to come up with an intelligent solution that takes into consideration where the soccer and athletic resources in this country lie and the ways we are fundamentally different from Europe
good post…I think you and Quit whining and Diego are actually all agreeing on most of this
How much less could you care? A lot less?
Irregardless, theirs a lot he's happy about to.
Jonathan Wallace
You mispelled irregardlessly.
You embiggened irregardless.
doughnuts, I didn't intend to genuflect on the rest of the soccer world. My point is the exact opposite. There are many in the US so brain washed to the idea that Europeans like Klinsmann know better about every aspect of soccer coaching/management that they support and make excuses for every hair-brained or self-serving thing he does. I agree, the rest of the world doesn't know much about soccer in the US but that is precisely my point. The one US player they did know and respect was Landon Donovan and they knew it was bone-headed to leave him off a roster of 23 American players.
Yes, most in the U.S. are so brainwashed they even believe Klinsmann is a good coach. Anyone who knows anything about soccer in Germany, and it is most of the population, knows Klinsi is a rotten coach. Just terrible.
Diego's Maradoughnuts
Good to know. Can you please ask them for their assessments of Bob Bradley, Bruce Arena, Steve Sampson, Bora Milutinovich, and Bob Gansler? These guys seem like a great resource for our program and our players. Thanks.
Diego's Maradoughnuts
Ok Inter. Sorry to come after you so hard.
In fairness, it's actually an important point for people to recognize that the informational advantage we have enjoyed over our more glamourous oppenents will be eroded as we contnue down the road we are on.
We have benefitted from our anonymity. Part of this can be attributed to laziness/arrogance, but plenty of it is simply due to time and access constraints. It doesn't matter if it's JK or Bradley or Arena who is sitting in the chair, the fact is opposing coaches have about 6 months after the draw to figure out exactly what the USA will look and play like. As a team we are frenetic in style depending on the personnel and opposition. Not much there to learn. As individuals, half our guys are unknown completely. They may know Dempsey, Howard or Donovan well enough, And they can crash-scout MLS tapes (pretty inefficient). They can calll clubs who have looked at purchasing them, but this data is limited and probably nonexistent for a guy like Besler or Beckerman. But we sometimes ambush teams because we are in a good position to do so. Portugal 2002 was our tour de force in this.
It will change and that's ok. Our players moving to Europe means recongition, which cuts both ways.
Gary Page
Those of us inside the US who really know soccer/football knew it was a mistake, too. And for those who disagree with this, can anyone find anywhere a coherent explanation from Klinsmann as to why he didn't choose Donovan? I never saw anything. And please, Brad Davis ahead of Donovan>?
Nate Dollars
he's got a contract through 2018, and ussf seems content with happened in brasil. i think he'll be alright.
wood chip zip
Any American coach would have been fired for the sum of what Klinsmann has done.
What's that? Completely re-tool a USMNT that was looking GRIM in 2011?
I'd remind people to take a look how bad we looked in 2011, when we got up two goals against Mexico in the Gold Cup…and then dropped four goals in machine-gun succession, reminding us all just how bad our backline looked.
As far as "core" guys, we had Altidore – who was struggling – Dempsey, Bradley, Howard, and Guzan…the other 18 spots in the 23 were pretty much question marks. Our backline was going to have to be completely re-tooled…Cherundolo was hurt by then (and was 33, I believe), we had Bornstein at LB, and Bocanegra and Gooch as our CB's, both on the wrong side of 30. Donovan decided to go take his sabbatical at that point; we had very little idea if he'd ever be back.
Things were looking BAD.
For whatever reason – and people don't make enough of this – there was a big talent gap between the Donovan/Dempsey/Howard generation and the guys who are now in the U23 age group. Right now it seems we have promising young players everywhere…but four years ago the cupboard was largely bare as far as young prospects went. Dempsey was born in 1983. If you look for prospects who were born 1984-1993…there just aren't many. Bedoya is OK; he plays in France. Zusi and Besler got some Euro offers. Omar Gonzalez could probably play in Europe. Bradley played for Roma but couldn't start…and kind of proved why in the World Cup. Geoff Cameron started for Stoke for a couple of years.
And that's it. There certainly wasn't a Dempsey or a Donovan in that age group…or, honestly, anything close.
But despite that, we're still loaded with promising young guys. And the younger generation of guys born post-1993 looks MUCH better. And now nobody's asking: "what happens after Donovan"…or "what happens after Dempsey or Howard". Because the emphasis has been: build the pool.
It astounds me that people can't see how much better the pool looks, right now. In that, Klinsmann has almost worked miracles.
Nate Dollars
would love to know specifically what 'miracles' klinsmann has worked.
pretty sure julian green came simply due to klinsmann–and that's very good–but i'm not sure how you can tell which of our up-and-comers were going to look promising whether klinsmann was here or not.
Klinsmann had almost nothing to do with the Julian Green situation. That's probably the reason he didn't play him until the last 15 minutes of the last game when there was not chance at victory.
Everybody was up in arms because of the "secret deal" he cooked up with Green to guarantee him a spot on the plane if only he'd commit to the USMNT. It's a disgrace, everyone said. He's a KID. He's not ready. Such deals are beneath us.
Now Klinsmann had nothing to do with Green being here, and didn't play him enough?
Make up your farkin' mind, peoples.
Wood Chip zip
Green pretty much admitted he chose the US because of Klinsmann.
Stop the Presses!
What else you got today, Dirk? I can't write this stuff down fast enough!
1. Pulling Run DMB off scrap heap, converting him to LB. Run DMB holds up entire World Cup despite being targeted by everyone.
2. Yedlin. Nobody thought he was ready. Klinsmann disagreed.
3. John Anthony Brooks. See #2. Especially since he was coming off a horrific showing against Ukraine.
4. Playing Cameron at CB when his club position was RB…and everybody (including me) was lobbying for him to play the same spot for the USMNT. Then moving Cameron to holding mid to stop Belgium from bullying us with Fellaini the way they had everyone else.
5. Kyle Beckerman. NOBODY wanted him on the USMNT. Klinsmann thought otherwise.
6. Jermaine Jones. Did ANYBODY think he was going to have as dominant a World Cup as he had? Lot of people were calling for him to be dropped outright.
I'll also add, in qualifying he got huge contributions from the likes of Eddie Johnson and Brek Shea, guys who have not exactly been easy for their clubs to handle.
This largely comes back to Klinsmann's decision to drop Donovan. People can't seem to get past it.
I myself, probably would not have done it…and yes, I think Donovan would have contributed a lot more than Wondo or Brad Davis ended up contributing.
That said…I also understand why Klinsmann did it…and in some ways, that decision did leave the USMNT in the clear position of NOT having to worry about replacing "legendary" players next cycle…because we already replaced them. And Germany and Belgium still would have been better than us, either way.
Beckerman was in the mix before with Bradley and I ALWAYS liked him…you use the word everyone and no one too liberally
Cameron was OK at CB, can't see any other evaluation of him considering his uneven play, some good and some not good
JJ? he was awesome before and had proven himself as under control years before this summer when he handled himself against our CONCACAF friends who baited him repeatedly–before Jurgen was coach
as for understanding Jurgen's move with LD as anything more than personal grudge, I think that's a bummer quozzel and better handled by calling a spade a spade–Jurgen screwed the pooch with that decision and his reasons (personal) magnify the blunder and arrogant manner in which he did it
Jurgen is media savvy. he knew the contingent of LD haters here and what LD stands for, which is American soccer made in America. For many, there is no way we can do it without leadership and guidance from abroad…it's soccer and we're just Americans! LD flipped the bird at that notion and some love it, others hate it
Nate Dollars
1. klinsmann did not convert dmb to left back. he's been playing there for years.
2. agreed.
3. plenty of people were ready to bring brooks, and some even wanted him to start the whole cup. i believe ives was one who did think that his selection was a surprise, but i'm not sure why.
4. cameron's best position is either CB or d-mid; doesn't take a genius to play him there.
5. that's a decent point, although it also displaced bradley from his best position.
6. like your point about brooks, calls for dropping jones were always ridiculous.
7 (or whatever). it's not a miracle to realize that you can get through qualification with some guys who might be struggling with their club teams. hell, every usmnt coach ever has had to do that.
Wood Chip zip
Miracles like the fact that the US had more muscle/overuse-related injuries than any other team in the World Cup?
That's a different conversation. Where is the line there?
We lost Altidore to a hammy. That might have been overtraining.
We lost Besler for one half – precautionary. Overtraining? Maybe.
Our fitness levels were also MUCH higher than anyone except Germany's. Once things settled down, Portugal couldn't run with us, and we took control of the game. They got a miracle at the end and it was a mental lapse, not a physical one. We were still coming at Germany at the end of that game despite them having a massive edge in possession. We were still coming – hard – at Belgium after 115 minutes while their players were dropping like flies.
You can argue Klinsmann made a huge mistake in not bringing a like-for-like replacement for Altidore (either Boyd or Eddie Johnson) and I wouldn't argue…and if you pointed a guy at Klinsmann's head, I don't think he'd argue either. That decision certainly bit us.
But "overtraining" them? Dunno. It's a fine line, granted…and when you push guys to the red line, they can indeed break…but you also want a team that can operate as close to the max as possible. Considering how many miles our guys were able to run, and considering that Germany won it all with the same approach, I…dunno.
One thing that does interest me is that new "Catapult" system that FSU employed for its football team…I'd like to see the USMNT employ it as well…it gives coaches a lot better idea of when to put the foot up the butt, and when to back off.
If we could have kept Altidore on the field, AND still had the fitness levels we had, it might have been a different World Cup.
Diego's Maradoughnuts
This is boring and repetitive. Why not say something new? Here is a suggestion….
One thing that I find striking about the dozen or so voices who have repeated the "Fire Klinsman" mantra several thousand times in this space over the last 3 months is that not one time… not ONE… have I heard anybody suggest a replacement or pitch a better option.
Is this not outright strange? If you don't like something, it seems like basic rhetorical strategy to try to convince people of a superior alternative. Something that might be agreeable to both of you.
Why not give it a shot? Because it's become predictable noise to hear the complaining that goes on every day in here. That's all it is… complaining– and to people who had zero to do wth the decision. We've heard it. We have documented your position. Plenty agree. Please advance the discussion
Tony in Quakeland
David Moyes, Tab Ramos, Jorge Pinto, Jose Pekerman for starters. Pekerman has a job at the moment, but we could have had him earlier. All four would be massively better choices.
We could have had Pekerman or Bielsa. And I'm pretty sure Ramos was handling the tactics in Brasil while Klinsmann was handling the subs.
Tony in Quakeland
I had the feeling Berti Vogts was handling tactics. The tactics didn't look much like what Ramos does with the Youth sides.
Wait, the US employed tactics in Brazil? Sorry missed that part.
Gary Page
Over the years I have seen a lot of speculation like this. It assumes that all these top foreign coaches are clamoring to get the US job. I have not seen any evidence of this. I remember a couple of cycles ago when the US was all hot to get Carlos Quiroz and made all sorts of offers and never could get him interested. Things may be changing, but for a long time the general perception was that the US is a soccer backwater. The bottom line is that we were lucky to get Klinsmann. I guess the fact that this is being debated at all is a good sign that more people are actually caring about this.
I don't know. I'd think that the US is one of the top ten jobs in the world at this point. Sure, you're not winning the World Cup, but unless you're Germany, Brazil or Argentina, Spain or Italy, you're not winning anyway. Compared to other teams of the same caliber, there's no pressure, you're going to make good money, live in Southern California, have a clear shot at qualification, and anything else is gravy. Where else are you going for that? Russia? Australia? Japan? Qatar? England is a fool's paradise, France is a total mess, Holland and Belgium are cheap and not going anywhere, Russia, well you have to live in Russia…Australia is never getting out of the group, and is way too far away, mexico? Croatia? Nigeria? C'mon. The US is definately at the top of the second tier of jobs.
Tony in Quakeland
And there has been plenty of talk of potential replacements. I have bought up Moves here several times. He has an affinity for American players and is great at organized a bunch of guys to play well against superior teams. (It's teams of superstars he doesn't handle.)
I bought up Pinto too. I would be over joyed if we looked as organized as Costa Rica did..
Moyes would have been great after Bob. It was around the time he'd coached Timmy and Landon at Everton.
Tony in Quakeland
Seriously. I can't think of a better choice.
Diego's Maradoughnuts
Not bad. I like this best of the ones I see here.
Doesn't exactly answer the question of "what do we do if we replace Klinsmann now", particularly as Bradley was "replaced by Klinsmann" more than he was "fired" in my view, based on what I've read about Gulati's rationale.
That aside, I don't know that Moyes is interested in taking on the new challenge of int'l management at the moment (for starters, I think Scotland would've hired him by now). He has a rep for being extremely hard-working and very hands-on, and at this point in his career it would likely be frustrating to be in a position where he gets so little time with his players.
But a good suggestion. He does know more than many about systematic develpment and implementation (this is what Gulati has really mandated JK to do). And you could say he has a better tacitcal resume, though I really believe choosing a coach based on tactical credibility in the current situation has very limited value comapred to our greater needs We'll be playing checkers against the top teams for years to come until we get a proper chess set.
All in all– I like Moyes as a name to keep in mind. How's his German?
Nate Dollars
if you haven't heard any suggestions, it's only because you're not listening.
i don't have a huge problem with klinsmann as manager (especially now that he's technical director, which is what he should've been in the first place), but i would've loved to get bielsa.
Diego's Maradoughnuts
This is very much the first I've heard, and I'm here pretty much every day. And I'll say it's a seroius breath of fresh air. I could actually listen to this all day by comparison.
Thanks guys. Keep it going. Let me think about the suggestions above and I will give you the respect of a thoughtful reply.
Nate Dollars
my dream team is still (as it was in 2010) bielsa as manager, klinsmann as technical director.
yeah, that's a huge amount of $$$, but i would love it. 🙂
Gary Page
Here are facts–the US national team had its best yearly record ever in 2013. The US qualified from the Hex with the highest point total in its history. Here's an opinion/fact–the US got out of what is arguably the toughest WC group it has ever been in. Try basing your opinions on facts. I thought it was a big mistake to leave Donovan off the WC roster, but do you really think any coach could have done better? Remember, Ghana beat us twice before, Portugal was rated 5th in the World and Germany won the WC.
Haven't you heard? Donovan got the call for the friendly in October. One can only hope that this will be the first of many call ups in t he near future for Landon.
Tony in Quakeland
In an alternate universe Klinsmann and Donovan had a conversation like this:
Donovan: This is it. I'm going to retire after this world cup
Klinsmann: Fantastic. Go out there and attack. Have fun. Leave them with more memories.
Donovan: Thanks. I will.
Thing is, is that really that farfetched? We would have gotten a Donovan like we're seeing now and like we saw in the last Gold Cup.
Would we have, though? Or is this newfound form Donovan's on a function of eminent retirement? Thierry Henry looks like he's pulling off a serious swan song as well.
Dunno. Klinsmann was the coach on the ground. He was the guy looking at these players every week, trying to assess who was on, who was off, trying to figure the best chemistry, the best mindset.
He made a very hard, controversial call…and given how Donovan looked in the Mexico friendly – which was, bad – and given Donovan's comments about how "he just couldn't train like he used to"…you can see how Klinsmann would have been supremely leery he was dealing with a blown player in Donovan.
Would Donovan have REALLY helped the USMNT advance past Belgium, or did we go as far as we were ever going to go, given the group we got, and the draw we got in the Round of 16?
Nobody gave us a chance of getting out of the Group of Death. We did it. We almost pulled a smash-and-grab against a very, very talented Belgium team.
That, to me, looks like about the max anybody could have ever asked for out of this USMNT squad…with or without Donovan.
So why so much hate?
if you don't think LD could have had a positive impact on that USMNT team, that's on you. would it have changed outcomes? we'll never know because Jurgen chose others who did what they did…that we know. could LD have helped vs. Belgium? look at who was on the field for us and answer that question honestly yourself
but as we all know we'll never know. that's what has many of us pissed…it only comes around every 4 years and we feel short changed
Diego's Maradoughnuts
And it's understood. Believe us, we have gotten the message and plenty sympathize. But nobody here made the decision, and none of us can help.
Discussing is good. But there's been nothing but retreads for at least a month. You know it's stale as well as I do bud. It just bums everybody out. The value of the discussion in resolving the disappointment has exhausted itself. Time to advance the ball… for everybody. Bring a new thought. Bring a superior solution. At least give everybody else something to work with
Diego's Maradoughnuts
I like the idea of Donovan initiating a discussion with Klinsmann… think it would have been ideal to do this earlier (as in before made a unilateral decision to step away from the program for an extended and likely needed break). Tim Howard did it this way and I that's what I think any of us would do when dealing with a boss.
LD made that decision…and then Jurgen INVITED him back. LD then played great…but the decision BEFORE being invited back and playing great is the determinant one? who cannot/could not let go? it's Jurgen, this is all his doing at this point. And many here defend it. weak
Diego's Maradoughnuts
I have a response, but I'm gonna leave it. There is only so much you can speculate about people's motives… we'll never really know and we've been on this forever.
See my comment above… Let's just shake hands and walk way from this one. Honorable men can differ. Nobody is leaving their position at this point. And I can think of 1,000 scenarios in which we are both way off base. For all we know, JK and LD were once tag team champions in an underground Mexican wresling federation, and this is just collateral damage somehow.
shake Diego
Go Galaxy!
Really? Cause he wasn't exactly dominating pre-World Cup.
Direct quote from Landon- "I realize very clearly that there's only so many more chances to do this, and the decision to retire and playing this way are intricately linked," Donovan said in a happy LA locker room. "That's given me freedom to relax and enjoy it, and it's taken all the pressure off, and that's when I play my best. It's been a lot of fun, I'm enjoying it, and most of all, I like winning."
I think he should have gone to Brazil regardless, but I don't think it would have let us progress any farther, and I don't think it was really that bad of a decision.
You do know that quote is in the article above, right? Wait, did anyone here even read the article? Or do we all just see any headline with Donovan's name in it and jump directly to the comments section to start the flame throwing?
Galaxy looked awesome last night. That third goal? Twenty six passes in the build up? Rumor has it that Barcelona has requested the tape on that.
So this turned into a Love/Hate Klinsmann thread but I want to hear who else saw Johnson's continuing classless behavior. First he claims a goal he didn't score, then he taunts the Galaxy fans when he's losing 3-1 and his team is getting waxed. Seriously, what happened to that guy?
Tony in Quakeland
He's kind of always been that way. Sometimes being an ass helps him succeed, other times it gets in the way. But ultimately it is the reason why he has never broken through to the next level, a level you'd think he's capable off.
By the way, thinking Klinsmann can't coach isn't hate. I think it's just the truth. Give him a figure head role recruiting players and talking up the game, fine. As a player, he was damn near my favorite. I don't hate him. I hate that he's coaching
hey Limey, cheers…I'll get over the LD thing eventually.
it's not love/hate altho I'm branded that. It's responsibility, and with it some humility, that our leader could exhibit, just come clean and admit making a mistake…would be true leadership and unite imo, would be a move that would earn him sincere respect from me, and from others, and hardly unprecedented for a coach…if we are one thing, it's fallible! But his fan base might not approve of that seems to me, already having justified and rationalized the LD cut and how it was done as sound
as for Johnson, it's part of his edge imo, for better or worse, either cuts for him or cuts himself but he's out there on it
I'm guessing you'll get over it before JK apologizes but I'm left with this feeling now that LD had a crappy 'I'm over it" attitude and that was cancerous to the squad. I think that's why he got left behind, he's just so darn emotional.
As for the love/hate i meant it more like a changing emotions thing not that anyone actually hates him, that would be a lttle extreme.
Brian S.
I could be looking through the wrong colored glasses but the way Donovan has been playing the past couple of months, he could be making a push for MVP.
…and no one east of Carson saw it,
Brian S.
The third goal was something special. 27 passes before tapping it in is, dare I say, Barcelona-esque.
Impressive string of passes, though Gordon was offside on the pass from Donovan. Doesn't take away from the focus of connecting that many passes for me, but I was struck by the announcers marveling at how wide open Gordon was, when I'm sitting there thinking "it's because he's offsides, stupid".
C'mon Chad no need to be bitter, I didn't think Gordon would do well either. BTW "Offsides?" Really? The term is offside and he wasn't.
Complete game, everyone was involved and its great to see. Donovan played with passion and it showed.
Brain Guy
Yes. According to the article title – Galaxy is currently heading to DC. Landon must be driving the pace car.
Dominating performance by LA, but what was the story with that PK call? It may have been the worst I've ever seen. I hope the league addresses it because it's embarrassing.
I wonder if Boswell had a fist full of shirt on the other side of Gonzalez from the camera – i.e. on the ref's side. Because it was hard to see much of anything wrong from the camera angles. My gut reaction was no way is that a penalty, but the ref's reaction was very definite and quick, which leaves me thinking there had to be something clear that wasn't on camera.
thought the same thing but can't see that angle
Gary Page
Bad call, but the worst ever? You need to watch more soccer. Here's one much worse–call against Gooch vs. Ghana in the 2006 World Cup made by referee Marcus Merk, whose name should forever be infamous for US soccer fans.
It was three nil at halftime, but when it became 3-1, it was dcu who was pushing to make it 3-2 and then the phantom pk made it 4-1. Closer than the score indicated. These teams could meet in the final and dcu will make it three and o against threir west coast rival. It would be right also with both teams sitting on four stars
LAG had eleven shots vs DCU's three. Eight of their eleven were on target. It looked more like LAG got up three points and relaxed. If "close" means LAG letting DCU run around with the ball after they were up three points then it was close. But if you mean DCU at any time in the game looked like they could win, then I disagree.
Dikranovich The penalty decision aside that scoreline was a fair reflection of the game. The Galaxy played the entire second half at around 60% effort.
who knows but could be many different players impacting a different game if all players play…DeLeon and Espindola for DC, Keane and Ishizaki for Galaxy, for example
Galaxy looking good, even without Keane last night. The passing and ball/player movement, quick and purposeful one and two touch passing, has been the difference since the SKC/MU games
The Galaxy seem to respond well to ass-whoopin's. They don't often lose badly, but when they do, it seems to snap them out of their malaise and put them back on track. I like that.
me too
Diego's Maradoughnuts
Now we're talking. There was a Galaxy game, wasn't there? And they looked super– seems they learned their lesson in Columbus, and filled their water bottles up on the way out with whatever they have been pumping through there.
Gary Page
When the Galaxy play like they did in the first half last night and first half against Vancouver, they are very hard to defeat. They play like Klinsmann wants the US team to play–quick one touch passing, good ball movement, good movement off the ball, the fullbacks joining the attack, and dominant ball possession. The first half I think Eddie Johnson only touched the ball a couple of times and probably 70% of the game was being played in the DC United half of the field.
Cliven Bundy
Someone will give Donovan the boots ,, its MLS
I'm so bored of this stupid Klinsmann thing, Donovan has moved on, so should we. I want to hear who else saw Johnson's continuing classless behavior. First he claims a goal he didn't score, then he taunts the Galaxy fans when he's losing 3-1 and his team is getting waxed. Seriously, what happened to that guy?
Seriously, people still arguing about the World Cup. It's over and done. I need to see EJ's taunt though lol. I hate DC, but he's still one of the most entertaining characters to watch.
4.07 in the highlights above. It's a close shot so you can't see who he is directing his 'words' to but it was to the Galaxy fans behind the goal.
+1, preach.
but look out, Jesse might come in here yelling at you for telling people how to think while simultaneously telling you how to think.
I noticed that too. He seems like a genuinely unhappy person.
Old School
In my expertise as a non psychiatrist its my professional opinion that an alarming amount of adults on SBI have panties in the butt-itis.
Its time to move on. You guys are worse than a crazy ex. Its beyond amusing and has become disturbing.
Gary Page
I think there are 2 major reasons for the continuing discussion. 1. Many people are concerned about Klinsmann's judgment and ability to coach and since he is our manager and general manager up through the next World Cup, then it is a relevant issue. Personally, I think it was a terrible mistake, but was balanced out by the good decisions he made. 2. People want to receive validation for their opinions that have been borne out by developments. Does anyone seriously think that Donovan would not have been an upgrade over both Davis and Wondolowski? Or how about Diskerud who played 0 minutes? I, like anyone, like to be proven right. It gives me and others credibility in the case of future opinions/predictions.
You are right! Jürgen's poor judgment and horrible man management led him to make a terrible mistake.


text:
The Underworld
The group entered the door and Mark's first instinct was to search for a light switch.
"Damn, no lights. How old is this little short cut anyway?" Mark asked.
"Hades didn't say, but it shouldn't be a long trip. It never is with Hades." Britney answered.
Mark turned on the flashlight to reveal an old hallway. It resembled old castle passages he'd seen in movies as a kid. The group moved foward into the dark until they came to a stair case.
"I thought you said this went across campus?" Oz said.
"It does," answered Chealsea, "but it goes into a basement first."
"Great," said Oz, "This is just how I wanted to spend my night, exploring some musty old passageways while students are turning up dead."
"Well we don't have much of a choice Oz. Lets get going." said Mark.
The four descended the stairs and came into what looked like an abandoned store room. There were some cleaning supplies and maintenance tools, as well as a gallon or two of gasoline. Cobwebs covered everything, and sections of the ceiling looked broken and there were cracks that let water flow into the room. This combined with the layers of dust covering the room created a smell that Mark imagined would be in a mausoleum. Just then Mark's heart nearly stopped.
The room was covered in bugs.
"Good job girls," came a voice from the dark.
Mark and Oz watched as Britany and Chealse walked foward to meet Hades as he emerged from the dark.
"It's time you boys stopped your little plan," Hades said, "This is my school now."
Mark noticed Hades skin was broken, but not bleeding. It was cracked all over, almost as if there was something trying to break out of the skin from inside. Sure enough Hades began to peel back his skin, revealing that he was a bug himself, and Mark was in his Hell as he watched Britany and Chealse do the same. It was all a trap, there was no secret passage. Just a hive of bed bugs and no escape.
"Lets get out of here Oz," Mark whispered.
But there was no answer. Mark turned to see his roommate in the grip of the exterminator.
"I thought you were an exterminator, kill these bugs not us!" Mark yelled.
The pale face of the exterminator stared back, and with a blank souless expression he answered, "I am an exterminator, and there is an infestation at this school. You humans have been on this campus far too long."
Mark watched as the exterminator began to shed his skin just like the others. In a panic Mark threw the flashlight at the exterminator who shrieked as it struck him in his face. Oz was now scrambling to his feet and the two darted across the room. The door was locked. This was the end. Mark and Oz were alone with the bugs. Mark had a lighter to see by, but they could only see the floor about a foot in front of them. All they had was each other, the flame of the lighter, and a canister of gasoline that lay on the floor at Mark's feet. He could hear the ticking of bug legs moving on every surface of the room. He had to act fast.

subset: commoncrawl


text:
Nevada coach Chris Ault has long emphasized the notion that every player on the team has to understand his role and succeed in that role in order for the team to have success. In that sense, the linemen must work as one cohesive unit. Skill players on offense have to be playmakers and game-changers. Sometimes defensive linemen have to take on more than one blocker in order for linebackers to make plays. And so on.
As part of that, each team must have its players who specialize in special teams. And that's why Nevada is fortunate to have a player like Andy McIntosh, a special teams star who is entering his fifth year in the program. McIntosh is listed as a wide receiver and he'll see some playing time as a pass-catcher.
"But his role is special teams and we are looking for him to be a leader on special teams," Nevada coach Chris Ault said. "Andy is one of those players who really understands and excels in his role."
But don't count out McIntosh, a three-time Academic All-WAC selection, contributing as a receiver. He made a pair of catches in Sunday's scrimmage, including one on a ball that was tipped at the line of scrimmage which forced McIntosh to adjust his body and make a tough catch while taking a strong hit from freshman Thaddeus Brown.
"He is the kind of guy that Nevada football is all about," Ault said.


text:
|Publication number
|Publication type
|Application number
|Publication date
|May 11, 2010
|Filing date
|Dec 14, 2005
|Priority date
|May 15, 2000
|Fee status
|Also published as
|Publication number
|Shuhei Yamamoto, Hiromi Yamashita, Masatoshi Kitagawa
|Original Assignee
|Hitachi, Ltd.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (26), Classifications (23), Legal Events (1)
|External Links: USPTO, USPTO Assignment, Espacenet
This is a continuation application of U.S. patent application Ser. No. 09/842,685, filed on Apr. 27, 2001, now U.S. Pat. No. 7,056,427 the entire disclosure of which is incorporated by reference herein.
1. Field of the Invention
The present invention relates to an electrophoresis apparatus which separates and analyzes samples such as DNA and protein by making use of a plurality of capillaries and a sample plate assembly used therefor.
2. Conventional Art
An application technology in which an array is constituted by combining a plurality of capillaries, an electrophoresis medium and a sample to be separated or analyzed are supplied to the respective capillaries and moved therethrough to thereby separate and analyze the object sample is well known, wherein a sample such as DNA and protein marked by a fluorescent material is supplied to the capillaries. Such application technology is, for example, disclosed in U.S. Pat. Nos. 5,366,608, 5,529,679, 5,516,409, 5,730,850, 5,790,727, 5,582,705, 5,439,578 and 5,274,240. In view of a through-put of the separation and analysis, it is much more advantageous to use electrophoresis with multi capillaries rather than electrophoresis with a flat plate gel.
A capillary array electrophoresis apparatus is basically constituted by such as a capillary array, an excitation light system including a laser beam source, a light receiving optical system which detects fluorescence and a voltage application unit which causes electrophoresis. In such capillary array electrophoresis apparatus the capillary array is constituted by aligning a plurality of capillaries in a plane shape, and a laser beam is irradiated to the capillaries which are filled with a sample (fluorescent sample) marked by a fluorescent material in parallel direction with the capillary aligning direction, then, through the lens action of the capillaries the laser beam is condensed and the laser beam is irradiated to the fluorescent sample in all of the capillaries when the laser beam is irradiated, the fluorescent sample emits fluorescence. Through detection by the light receiving optical system of the fluorescence emitted from the fluorescent sample in a direction substantially perpendicular to the laser beam irradiation direction, the measurement of the sample is performed.
In order to automatically supply at least a sample to capillaries in a multi capillary array, a sampling device is used. An object of the present invention is to provide an electrophoresis apparatus which can perform separation and analysis of samples by supplying many samples continuously to the capillaries and a sample plate assembly used for the apparatus.
In an electrophoresis apparatus in which a sample marked by fluorescent material is separated in capillaries through electrophoresis by making use of a capillary array formed by a plurality of capillaries, fluorescence emission is caused by irradiation light to the sample and through detection of the fluorescence the sample is analyzed, the present invention provides an electrophoresis apparatus which comprises a sample plate assembly including two sets of a tray at a sample supply portion of the capillary array, an adapter for the tray, a sample plate mounted on the adapter, a septum mounted on the sample plate and a septum cover mounted on the septum, wherein two sample plates include respectively different numbers of wells, and the tray, the adapter, the septum and the septum holder in each set being adapted to the corresponding different sample plate. Further, the tray includes a sensor for discriminating the adapter, and thereby the sample plate assembly is correctly placed on the tray. Still further, the tray is designed to permit mounting a plurality of units thereon according to the sample plate assembly being mounted thereon and the respective trays are provided with a sensor which permits to judge whether the unit is correctly mounted.
Further, the present invention provides a sample plate assembly including two sets of a tray at a sample supply portion of the capillary array, an adapter for the tray, a sample plate mounted on the adapter, a septum mounted on the sample plate and a septum cover mounted on the septum, wherein two sample plates include respectively different numbers of wells, and the tray, the adapter, the septum and the septum holder in each set being adapted to the corresponding different sample plate.
An electrophoresis apparatus according to the present invention will be explained with reference to
As shown in
A gel supply side 305 of the capillary array 301 is connected to a gel pump system 304 and the gel is supplied from a reservoir 312 by a syringe 310. Another electrode 307 is attached to the reservoir 312. A detection portion 308 of fluorescence generated from a sample marked by fluorescent material which is separated by electrophoresis in the capillaries is disposed outside the space in the thermostatic oven 311.
An entire operation of an electrophoresis apparatus using a capillary array according to the present invention will be explained with reference to
As shown in
Now, the operation of the capillary array electrophoresis apparatus will be explained. As shown in
Now, the operation sequence of the capillary array electrophoresis apparatus will be explained. The buffer liquid 436 contained in the buffer liquid container 417 is injected into the capillaries from the buffer liquid injection port 430. Then, the conductive fluorescent sample injection port 432 is immersed into the fluorescent sample container 418 filled with the fluorescent sample 434 to inject the fluorescent sample 434 into the capillaries. Thereafter, the conductive fluorescent sample injection port 432 is immersed into another buffer liquid container (not shown) containing a buffer liquid and a high voltage is applied between the buffer liquid injection port 430 and the fluorescent sample injection port 432 from the high voltage power source 419 to cause electrophoresis in the capillaries. Since the moving speed in the electrophoresis is proportional to the magnitude of electric charge of the molecules and is antiproportional to the size of the molecules, the fluorescent sample 434 is separated. Through the continuous application of the high voltage for a long time the electrophoresis is caused for a long time and the fluorescence 435 emitted during the electrophoresis is continuously measured.
The sample introduction portion 432 is structured by inserting capillaries into stainless tubes. Respective stainless tubes are soldered to an electrode plate with a protective cover and through application of a voltage to the connecting portion 431, the voltage is applied all of the stainless tubes. As has been explained, since the capillary array itself is provided with all necessary functions including the buffer liquid injection port 430 attached to the buffer liquid container 417, a light detection portion 429 in which laser beam is irradiated and the fluorescence is detected and the sample introduction portion 432 through which the fluorescent sample 434 is introduced and a voltage necessary for electrophoresis is applied thereto, when an exchange of the capillary array is required, the capillary array can be exchanged with a very easy handling.
Further, the top of the fluorescent sample injection port 432 is sealed by an adhesive so as to prevent carry over of such as the sample. A kind of the adhesive used is an epoxy series adhesive and the same is fully cured so as not to affect the electrophoresis. Gaps between capillaries and insertion portions therefor in the sample introduction portion 432 and between the fluorescent sample injection port 432 and the protective covers are sealed with an adhesive. Thereby, a possible electric insulation reduction is prevented which can be caused when water contained in the sample and the buffer liquid penetrates into the covers of the stainless tubes.
When once detaching the capillary array from the apparatus and storing the same after measurement of the sample, a dry preventive container cover (not shown) is attached so as not to dry the buffer liquid 436. The container cover is a dry preventive cover for the sample introduction portion 432. The container cover is attached to the sample introduction portion 432 while charging pure water therein. The container cover is provided with an O ring to thereby prevent a possible drying. It is also effective to provide a dry protective cap (not shown) for the buffer liquid injection port 430. In such instance the cap is set onto the buffer liquid injection port 430 under the condition in which a small amount of pure water is likely filled therein. When the inner diameter of the cap is determined to be smaller than outer diameter of the buffer liquid injection port 430 by about 5-15% to thereby prevent a possible drying. As a material of the cap, silicon rubber is preferable, because the silicon rubber causes no adverse effect to the buffer liquid and the electrophoresis. These cover and cap also work to protect the top end thereof and to prevent contamination thereof, when shipping the capillary array to a customer.
Each of the capillaries used in the capillary array as explained above is a fused quartz tube having inner diameter of 50�10 μm and outer diameter of 340�20 μm. Since the fused quartz tube itself breaks very easily, a polyimide coating having thickness of 15�5μm is applied on the surface of the capillary. In view of limiting amount of fluorescent sample 434 it is desirable to reduce the inner diameter of the capillary, however, on the other hand in view of a concave lens effect due to refractive index difference between the fluorescent sample 434 and fused quartz, the capillary having a too small inner diameter makes the measurement difficult. Therefore, the inner diameter of 50˜100 μm is preferable for the fused quartz tube. Further, in order to suppress the above refractive index difference it is preferable that the outer diameter of the fused quartz tube is small, however, a too small outer diameter makes assembly thereof difficult because of static electricity, therefore, the outer diameter of 250˜350 μm is preferable for the fused quartz tube. The coating material for the capillary is not limited to the polyimide, a material having an equivalent electrical insulation and other properties as those of polyimide can be used.
A sample plate assembly used for the auto sampler according to the present invention is formed into a four layer structure by laminating a plate adapter, a plate, a septum and a septum holder. The above unit is mounted on a tray of the electrophoresis apparatus according to the present invention. In order to reduce load for a person performing analysis, the analysis is performed by charging a sample to be analyzed into a marketed microtiter plate which is marketed from many manufacturers. These microtiter plates respectively include the corresponding adapters and any type of microtiter plates can be attached to an auto sampler. These microtiter plates can be classified into two kinds depending on the number of samples which can be charged into one plate.
One of the microtiter plates includes wells of 8�12=96 holes for introducing and holding samples into the plate and another microfiter plates includes wells of 16�24=384 holes. Sample plates referred to hereinbelow indicate these plates and the former sample plate will be referred to as a 96 sample plate and the latter as a 384 sample plate. Since in the electrophoresis apparatus according to the present invention two sets of the above sample plates can be mounted at the same time, a continuous analysis of 384�2=768 samples in maximum can be performed.
The septum 2 has a function of preventing evaporation of the samples and further serves to function of wiping off such as sample deposited at the top end of the capillaries. In the electrophoresis apparatus according to the present invention, a number of analyses per day for a standard sequence analysis is set to be 96 samples, therefore, with the present apparatus which permits mounting of two 96 hole sample plates at the same time, an automatic operation with no person can be performed in two days. Therefore, the evaporation of such as the sample and buffer solution is an important problem. Through fitting the septum 2 onto the sample plate 3 the evaporation of such as samples is prevented.
Further, the top end portion of the capillary array performing electrophoresis is immersed during the sample analysis into the sample, buffer liquid and water depending on purposes of analysis. The gel causing the sample in the capillaries electrophoresis is exchanged for every electrophoresis, therefore, in such instance the top ends of the capillaries are immersed in a waste liquid vessel (water) in order to receive the exhaust gel from the top ends of the capillaries. Thereafter, in order to introduce the sample into the capillaries, the end terminals thereof together with the electrode provided at the top end of the capillary array are inserted into the sample, and a high voltage is applied to the electrode and the sample is introduced into the respective capillaries. Further, when performing electrophoresis, the top ends of the capillary array are immersed into the buffer vessel so as to prevent electrical discharge during the high voltage application as well as to cause the sample electrophoresis.
When the sample plate assembly moves between the waste liquid vessel, the sample and the buffer vessel, the top ends of capillaries are immersed once or a plurality of times in a cleaning vessel (water) for cleaning so as to prevent mixing of such as sample and buffer liquid deposited at the top ends of the capillaries into other solution. The septum 2 is made of silicon rubber and is provided with projections so that the top ends thereof are inserted into respective wells in the sample plate 3, and cuttings are provided at the top ends thereof so as to permit passing of the capillary top ends therethrough. When the auto sampler moves, the capillaries once and always lower to a height so as not to touch the auto sampler, therefore, every time when the capillaries lower, water, sample and buffer liquid deposited at the top ends of the capillaries are wiped off by these cuttings, thereby, mixture of these to other solution is suppressed in minimum.
The septum holder 1 is a holder for securing the well plate and septum to the adapter.
Now, the structure of the septum holder (positioning guide) will be explained.
In order to reduce time for a person performing analysis, the septum holder 1 is configured in a simple structure, in that the septum holder 1 can be secured to the adapter 4 only by fitting to two sets of pawls 10 provided at respective right and left sides to the flanges 9 of the adapter 4. Therefore, it is possible that the septum holder 1 can be fitted to the plate adapter 4, while being offset in longitudinal direction. If the septum holder 1 is erroneously secured to the plate adapter 4, the holes in the septum holder 1 through which capillaries pass do not align the wells in the sample plate 3 which breaks the capillaries. In order to prevent such possibility, there are provided projections 6 at the portions where the four pawls 10 are formed at the side of the septum holder 1 and if the projections 6 do not match with guides 7 at the side of the plate adapter 4, the pawls 10 can not fit to the flange 9 and the septum holder 1 can not be attached to the plate adapter 4. With this measure, a person performing analysis can attach the septum holder 1 to the plate adapter 4 without special attention, and further an attachment to an erroneous position never occurs.
The auto sampler according to the present invention can be adapted to different plates (having different height) by making use of the adapter. Among the sample plates used in the electrophoresis apparatus according to the present invention, even with sample plates having the same 96 wells a plurality of types are marketed, and such as the shape, size and well depth thereof are different depending on the manufacturers.
For example, when the minimum sample amount for the electrophoresis apparatus according to the present invention is set at 10 μl, the height from the bottom of the well to the sample liquid surface for one of marketed 96 sample plates is about 2.5 mm, therefore, in order to surely introduce the sample into the capillary, the capillary has to be inserted to the height of about 1 mm from the bottom of the well. However, the height of the well bottom greatly varies depending on the sample plates marketed, therefore, if the sample plates are used without careful discrimination, there arises a problem that the top end of the capillary may touch the bottom of the well to break the capillaries or may not reach the sample to thereby fail the introduction of the sample into the capillary.
Therefore, in order to correct such-as the bottom height and the position of center axis of the well in the sample plates of respective manufacturers, plate adapters corresponding to respective sample plates are prepared and are used in combination with the respective sample plates, therefore, the center axis and the bottom height of the well for any sample plates having 96 wells are set equal with respect to tray 5 in the auto sampler. Thus, it is sufficient if the auto sampler only discriminates number of wells in that 96 or 384, of the sample plate assembly mounted thereon and determines control method depending on the discrimination, the auto sampler can always be moved for any type of 96 sample plates mounted under the same control method.
Further, it is difficult to separate at a glance the 96 well sample plates of respective manufacturers depending on the configurations thereof, therefore, in order not to be attached to a wrong plate adapter or in a wrong direction, a measure is taken for the respective plate adapters. For example, in the case of the sample plate 3 as shown in
The auto sampler according to the present invention can be adapted to 96/384 well sample plates with an identical structure. Any two sets of the plate adapters for 96 well sample plate and the plate adapters for 384 well sample plate can be attached to the tray 5 as shown in
The electrophoresis apparatus according to the present invention can automatically discriminate the 96 plate and the 384 plate. As illustrated in
The positional relationship between the light shielding plate 8 and the first cut-out 11 is inverted with respect to the plate adapter for 96 well sample plate and the plate adapter for 384 well sample plate, therefore, when the plate adapter 4 is attached to the tray 5 and signals from the two photo diodes 13 are compared, it is possible to discriminate whether the attached plate adapter is for the 96 well sample plate or for the 384 well sample plate. Further, with a second cut-out 12 as shown in
The auto sampler according to the present invention is provided with a stopper plate having a structure including an electrical discharge preventing measure.
For preventing the above, in the present apparatus a stopper plate 14 as shown in
Now, when the, capillary array is inserted into such as the sample plate and the buffer vessel, the stopper plate 14 is shifted upward due to the upward movement of the auto sampler, therefore, the stopper plate 14 pushes down the sample plate assembly and the buffer vessel by a larger spring force than the friction force. Thereby, when the auto sampler starts the downward movement, the stopper plate 14 always pushes down the sample plate assembly and the buffer vessel, there are no chances that such as the septum remains at the capillary array.
Further, the bottom face of the stopper plate 14, namely the face which contacts to such as sample plate assembly and the buffer vessel is structured not to be flat but uneven as shown in a cross section in
The auto sampler for the electrophoresis according to the present invention does not drop even when the power source is turned off, but can be pushed down manually. During analysis of a sample when the gel in the capillaries is dried, the introduction of the sample and the electrophoresis are disturbed. However, the top end portion of the capillary array which is subjected to the fear of drying is immersed in a liquid such as the sample and the buffer liquid substantially all the time during the analysis, although during the movement of the auto sampler between wells the top ends of the capillaries are exposed to the outer atmosphere, the exposed time is very short such as a few seconds, the gel in the capillaries does not generally dry during an analysis. Further, after completing an analysis or when interrupting analysis because of error detection, the top ends of the capillaries wait while being immersed in the buffer liquid, therefore, the gel at the top ends of the capillaries never dries before the subsequent analysis.
For the drive of the auto sampler, a drive scheme in combination of a stepping motor and a ball screw is basically employed, and during the time when the power source is made for the apparatus the stepping motor is always placed under an excitation condition, therefore, after completing an analysis, a condition that the capillaries are inserted into the buffer vessel, in other words the condition in which the auto sampler is shifted upward, can be maintained. However, sometimes it is desired to break the power source of the apparatus, because it is presumed to be long to the subsequent analysis or a person performing the analysis wants to interrupt the power source of the apparatus during an analysis for some reason, it is necessary that the auto sampler holds the position (the height in Z direction) before the power source is interrupted even under a condition where the power source of the apparatus is not made, in other words under the condition that the stepping motor is not excited.
On the other hand, when it is desired to remove the capillaries from the apparatus after the power source is interrupted, it is difficult to remove the capillaries if the capillaries are kept at the position where the capillaries are immersed in the buffer liquid, the auto sampler has to be pushed down manually to the height which permits the removal of the capillary array. Further, when the auto sampler can not operate normally because of occurrence of an abnormality, it is conceived necessary that the auto sampler is moved manually after interrupting the power source of the apparatus. For this reason, after the power source of the apparatus is interrupted, namely even under the condition that the stepping motor is not excited, the auto sampler always has to be kept at a predetermined position and if required the auto sampler can be moved manually by a person himself performing the analysis or by a person himself performing maintenance and inspection.
Fa=W�sin(arctan(P B /D B�π))[N] (1)
and the total friction force F[N] acting to the table can be expressed as follows;
F=−μ 0 �W�cos(arctan(P B /D B�π))[N] (2)
Accordingly, a condition where the table 20 and the work never drop downward even when the stepping motor is in a non-excited condition is as follows;
|Fa|<|F| (3)
Now, as an inherent force which works so as not to drop the table, there is, for example, a detent torque (static torque at the time of no excitation) of the stepping motor, however, such torque is very small with respect to the forces now concerned and no substantial contribution to the above inequation (3) is considered, therefore, the inclusion thereof is omitted. In the present auto sampler, since |Fa|≈11.4[N], the above inequation is satisfied, therefore, the auto sampler never drops even under the condition that the stepping motor is placed under no excitation condition. Further, if the auto sampler is pushed downward with a force more than |F|−|Fa|≈7[N], the auto sampler can be easily pushed down. The relationship with regard to magnitudes of the forces expressed by the equations (1) and (2) can be determined by the pitch PB of the ball screw and the shaft diameter DB of the ball screw. The shaft diameter DB of the ball screw is determined in advance in view of the structure of the auto sampler and the required mechanical strength thereof so that alternation thereof is difficult, however, the pitch PB of the ball screw can be selected with comparatively a large tolerance if such as drive speed of the auto sampler and the operating condition of the stepping motor are properly adjusted, therefore, the inequation (3) can be realized easily.
The auto sampler according to the present invention makes use of a position guide (reference surface) at the time of assembly. When 384 well sample plate is used, the diameter of capillary insertion port in the sample plate assembly minimizes and the inner diameter of the septum holder in this instance is 2.3 mm. Since the outer diameter of the capillary including the electrode thereof is 0.71 mm, in order that a single capillary can be inserted into a well without touching the inner wall of the septum holder, the distance between the center axis of the capillary and the center axis of the well in the X Y plane is required to be less than about 0.8 mm.
Further, in the present apparatus which can perform a continuous measurement to the maximum of 786 samples, the auto sampler moves to sample positions in the maximum of 48 points, therefore, if the orthogonality of X and Y axes of the auto sampler offsets, there arises a problem that even when the capillaries can be inserted into the wells without troubles at a certain sample position, the capillaries touch to the septum holder at another sample position.
In view of the above problem, a high orthogonality with regard to X, Y and Z axes is required at the time of assembly of the auto sampler, therefore, in order to reduce time for the assembly as much as possible, all of the parts relating to the orthogonality with regard to X, Y and Z axes are provided with reference planes. Therefore, it is simply enough to assemble the same while matching the respective reference planes with each other. Thereby, the time required for the assembly and adjustment thereafter is greatly reduced, and further, such as parts exchange during maintenance can be performed easily.
In the electrophoresis apparatus according to the present invention positioning pins are provided for the auto sampler and the oven unit, a system is constituted which automatically reproduces their relative positions during the exchange of the unit.
The orthogonality of the auto sampler with regard to X, Y and Z axes has already been explained, the positional accuracy required for the present apparatus is further complex which uses a capillary array constituted by 16 pieces of capillaries. The capillaries are aligned in a grid shape of 2�8 at the sample introduction portion of the capillary array, therefore, even if the orthogonality of the auto sampler with regard to X, Y and Z axes is highly accurate, if the grid axes of the capillaries aligned in a grid shape are not in parallel with the X, Y and Z axes, it is not certain that all of the 16 pieces of capillaries can be inserted into the corresponding wells without trouble.
Although the sample introduction side of the capillary array is attached to the oven unit, if the capillary array is always correctly attached to the oven unit, the positional relationship between the capillary array and the auto sampler is determined by the positional relationship between the oven unit and the auto sampler. The positional relationship between the oven unit and the auto sampler can be satisfactorily performed by adjusting the respective relative positional relationship during assembling the both into the box body of the apparatus, however, only with such adjustment, once one of the units is detached from the box body of the apparatus for such as repairment and inspection, the prior positional relationship of both is destroyed and in order to reproduce the original positional relationship another readjustment is necessitated.
Therefore, two pieces of positioning pins for respective units are provided at the box body of the apparatus, and for both units corresponding guides which permit insertion of the respective positioning pins are provided. The positional relationship of these positioning pins is adjusted in advance in the box body of the apparatus and once such adjustment is performed, no readjustment thereof is required. Thus, even when one of the both units or the both units are detached from the box body of the apparatus, when attaching the same subsequently, if the units are attached along the positioning pins provided at the box body of the apparatus, the positional relationship of the both units can be reproduced without adjustment.
Now, a structure/jig (target, ruler, etc) of a calibration use device in the electrophoresis apparatus according to the present invention will be explained.
The auto sampler control unit which performs control for the auto sampler stores positional information (in X, Y and Z coordinate system) common between devices for moving the auto sampler and inserting the capillaries into such as the sampler plate and the buffer vessel. However, in an actual apparatus, the relative distance between the auto sampler and the capillary array finely varies device by device, therefore, the control unit performs correcting calculation of the positional information based on calibration value (offset value) stored for every device, and thereafter drives the auto sampler by making use of the coordinate system after the correction. The present apparatus is provided with a calibration function which determines the above calibration value, and the calibration value is stored for every apparatus in a built-in control PC for the apparatus while permitting rewriting thereof.
In the calibration of the auto sampler, a target 21 in a frust conical shape provided on the tray 5 in the auto sampler is determined as a reference position in X Y direction and a line 22 indicating the liquid surface in the buffer vessel to be attached to the tray 5 is determined as a reference position in Z direction, the auto sampler is moved to a position where the top ends of the capillaries coincide with these positions and the X, Y and Z coordinates of the auto sampler at this moment are stored as the calibration values. The coordinates of these reference positions are in advance stored in the auto sampler control unit and the positional coordinates of such as respective wells in the sample plate and the buffer vessel likely stored are determined based on the reference coordinates. Accordingly, through determination of difference between the calibration values determined by the calibration of the auto sampler and the coordinates of the reference positions for the calibration which are stored in advance in the control unit, the coordinates such as respective wells in the sample plate can be corrected.
Hereinbelow, the above calibration function will be explained in detail.
Further, for the calibration in Z direction, the target line 22 (a line indicating amount of buffer liquid) printed on the buffer vessel as illustrated in
When the calibration of the auto sampler is performed in the manner as explained and the direction of the capillary array and the auto sampler is correctly adjusted, the capillary array correctly moves to all of the positions on the tray in the auto sampler. However, such is based on a precondition that 16 pieces of capillaries in the capillary array are correctly aligned in 2�8 matrix. Namely, the calibration of the auto sampler is performed by selecting one capillary among 16 pieces of capillaries as a representative, the remaining 15 pieces of capillaries have to be correctly aligned with respect to the one representative capillaries. However, it is possible that a person performing an analysis can by mistake bend a capillary during handling the capillary array, in such instance, it is indefinite whether all of the 16 pieces of the capillaries can be normally inserted into the corresponding wells in the sampler plate.
Therefore, another function for checking whether the capillary which was bent by mistake by the person performing the analysis can be used as it is and whether the manually straightened capillary by the person can be used is necessitated. For this purpose, the present apparatus is further provided with an array checking function which performs the above checking. When performing the array checking function, a capillary array of which alignment is required to be checked is attached to the apparatus and after performing calibration of the auto sampler depending on necessity, the array target 23 for checking the alignment is attached on the buffer vessel as illustrated in
A column 24 serves to lift the stopper plate (as shown in
According to the present invention, many number of samples can be automatically supplied to the capillaries.
|Cited Patent
|Filing date
|Publication date
|Feb 24, 1992
|Dec 28, 1993
|The Regents Of The University Of California
|Capillary array confocal fluorescence scanner and method
|Oct 12, 1993
|Nov 22, 1994
|Hitachi, Ltd.
|Electrophoresis gel migration apparatus
|Mar 13, 1992
|Jan 24, 1995
|Applied Biosystems, Inc.
|Capillary electrophoresis
|Jun 3, 1993
|Aug 8, 1995
|The Governors Of The University Of Alberta
|Multiple capillary biochemical analyzer
|Jan 13, 1995
|May 14, 1996
|Hitachi, Ltd.
|DNA detector and DNA detection method
|Nov 7, 1994
|Jun 25, 1996
|Hitachi, Ltd.
|DNA detector and DNA detection method
|May 19, 1995
|Dec 10, 1996
|Iowa State University Research Foundation, Inc.
|Multiplexed capillary electrophoresis system
|Jan 19, 1996
|Mar 24, 1998
|Hitachi, Ltd.
|Capillary array electrophoresis system
|Feb 5, 1997
|Aug 4, 1998
|Brookhaven Science Associates Llc
|Laser illumination of multiple capillaries that form a waveguide
|Jul 7, 1998
|Jun 29, 1999
|Spectrumedix Corporation
|Automated system for multi-capillary electrophoresis having a two-dimensional array of capillary ends
|Feb 20, 1998
|Jan 25, 2000
|Hitachi, Ltd.
|Fluorescence detection capillary array electrophoresis analyzer
|Feb 12, 1999
|Mar 20, 2001
|Perkinelmer Instruments Llc
|Autosampler having gripper with shape memory metal actuator
|May 16, 1995
|Jul 10, 2001
|Ashok Ramesh Sanadi
|Method and apparatus for preventing cross-contamination of multi-well test plates
|Aug 11, 1999
|Apr 2, 2002
|Spectrumedix Corporation
|Motorized positioning apparatus having coaxial carrousels
|May 27, 1999
|Nov 26, 2002
|Orchid Biosciences, Inc.
|Multiple fluid sample processor and system
|Aug 19, 1999
|Dec 17, 2002
|Caliper Technologies Corp.
|Indicator components for microfluidic systems
|Apr 25, 2001
|Jun 24, 2008
|Hitachi, Ltd.
|Capillary array electrophoretic device and auto sampler used for the device
|Jul 10, 1990
|Jun 12, 1991
|Schering Corporation
|Gas chromatograph/autosampler coupling assembly
|Mar 6, 1998
|Sep 16, 1998
|Japan Science and Technology Corporation
|Sample plate and multicapillary electrophoresis apparatus
|Title not available
|Title not available
|Title not available
|Title not available
|May 14, 1998
|Nov 19, 1998
|Aurora Biosciences Corporation
|Automated robotic system and method for rapidly screening and identifying useful chemicals in liquid samples
|Feb 19, 1999
|Aug 26, 1999
|Tetragen, S.A.
|Capillary electrophoresis apparatus and method
|Sep 8, 1999
|Mar 23, 2000
|The Perkin-Elmer Corporation
|Sample handling system for a multi-channel capillary electrophoresis device
|U.S. Classification
|204/601, 204/194, 422/64, 204/605, 204/453, 204/603, 204/604, 204/452, 204/451, 204/455, 204/193, 422/63
|International Classification
|Cooperative Classification
|European Classification


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New Braunfels Herald Zeitung (Newspaper) - October 8, 2003, New Braunfels, Texas WEDNESDAY, OCTOBER 8, 2003 RAVE REVIEWS Glenn Bishop, lead cook at Eden Home, gets rave reviews from the people who rely on him to provide their meals. Fags IB SPORTS UNICORNS UP Cross-town rivalry gets real Tuesday night as the Unicorns square off with Canyon and come out ahead. Page BA Serving New Braunfels and Comal County since 1852. Vol. 152, No. 281 16 pages, 2 sections CLICK 500 herald-zeitung.com 56825 00001 ly cloudy High Low I 84 68 1 • Details .... 1B I DEAR ABBY 4B j CLASSIFIEDS 6-8B I COMICS 3B I CROSSWORD 3B | FORUM 4A j APPLAUSE 5-6B I SPORTS 6-7A I TV GRIDS 4B SpRi 'Governator' terminates Davis See Page 3A Cub Scout TJ. DeWaters proudly displays the awards he received during a ceremony Tuesday night. See SCOUTS Page 3A By Erica Warner Associated Press Writer LOS ANGELES — Californians banished Gov. Gray Davis just 11 months into his second term and overwhelmingly elected action hero Arnold Schwarzenegger to replace him Tuesday—a Hollywood ending to one of the most extraordinary political melodramas in the nation's history. Voters traded a career Democratic politician who became one of the state's most despised chief exec utives for a moderate Republican megastar who had never before run for office. Davis became the first California governor pried from office and only the second nationwide to be recalled. "Tonight, the voters did decide it's time for someone else to serve, and I accept their judgment," Davis said in conceding. He pledged to work for a smooth transition. ican paradise — a state mired in economic troubles, awash with deficits, now governed by a Republican chief executive with no political experience and a Democratic legislature. Partial returns showed the recall favored by Arnold 1,448,449 voters, or 55.9 Schwarzenegger percent, and opposed by 1,143,613, or 44.1 percent. Other returns had Schwarzeneg- To the victor goes a spoiled Amer- ger ahead with 1,201,365 votes; Democratic Lt. Gov. Cruz Bustamante with 739,367; Republican state Sen. Tom McClintock with 303,043; and Green Party candidate Peter Camejo with 50,182. "This is a great day for California. ... In response to a common danger, the people of California rose to their duties and ordered a new direction for our state," McClintock said in conceding. Schwarzenegger prevailed despite a flurry of negative publicity in the campaigns final days, surviving allegations that he had groped women. DWI court frenzy was false alarm FRONTand Center Scouting illustrates character standards This story is the third in a three-\xiri series examining initiatives designed to help children develop good character. By Dylan Jim6nez Staff Writer Eduardo Segura, ll, smiled from ear to ear when he received his second Webe-lo badge in a Boy Scout advancement ceremony Tuesday night. He received a blue mark and a feather necklace. "Whoa!" the crowd of more than 70 Cub Scouts said. The Boy Scouts are an organization aimed at teaching character and values to youth, and, like character-based initiatives in schools and churches, the scouts program influences the lives of thousands of youth. like other out-of-school programs such as 4-11, which emphasizes development of "head, head, hands mid health," scouting illustrates extracurricular character learning and involves 1,609 youth and 600 adults in the Comal County program The Alamo Area Council of Boy Scouts mission is "to prepare young people to make ethical choices over their lifetimes.' That mission is accomplished bv reinforcing the Scout Oath and Scout Law. Scouts take an oath to help others, remain moral and perform duties to God and country. Their values are trustworthiness, loyalty, helpfulness, friendliness, courteousness, kindness, obedience, cheerfulness, thriftiness, courage, Livestock association set to showcase building plans See plans for the new show and meeting facilities I 3 to 6 p.m. Saturday I By the Extension Office, off Texas 46 at Resource Drive By Ron Maloney Staff Writer The Comal County junior Livestock Association Saturday will unveil to the public plans for the new show and meeting facility it hopes to build within the next five years. Plans include a bam, arena and meeting facility, parking for about 800 vehi cles and pens for student livestock projects. The county bought the property earlier this year, declared it a county park and leased half of it to the livestock association for $1 per year for 50 years. When the association builds a facility, it will move the annual junior livestock show from the Comal County Fairgrounds. In addition to the annual livestock show, the building could be used for other events, association President David Waldrip said. "It's certainly going to be something the people in the county can utilize and have fun with," Waldrip said. "It's going to be a versatile building." Livestock association secretary Liz Mooney said a drawing of the facility would be displayed Saturday, along with floor plans. "We're going to stake out where we hope to put it so people can get a visual on where it's going to be," Mooney said. The public is invited, Mooney said. "We're going to show people what we've got and try to get them fired up," Mooney said. Teach me Read about a teacher who volunteers her time teaching English at § second language. By Ron Malonoy Staff Writer Some suspected drunken drivers' late-August rush to the courthouse might have been for naught because new drunken driving penalties likely will take effect with offenses — not convictions — that occurred alter Sept. I. Gov. Rick Perry is expected to sign legislation enacting strict new drunken driving penalties — several weeks after a run on courts by accused drunken drivers seeking to resolve their cases before the law took effect. Under the new law, those convicted of drunken driving are subject to $1,000 annual fees for three years after conviction. The annual surcharge runs $2,000 for convictions with a blood-alcohol count of 0.16, which is double the legal limit. In August, District Attorney Dib Waldrip's office worked overtime processing drunken driving cases for defendants who chose to plead "no contest" or "guilty" before the law took effect Sept. I. County Court-at-Law Judge Brenda Chapman put on extra court dockets to accommodate accused drunken drivers who wished to resolve their cases before the deadline. The typical caseload for DWI cases doubled in August as local officials worked to resolve outstanding cases, said Bobbie Koepp, who supervises the County Court-at-Law clerical staff for County Clerk Joy Streater. "Judge Chapman set aside the Friday before the deadline for DWI cases," she said. "I think we took 30-something pleas that day alone." Heavy dockets are not unusual at county court, Koepp said. On Aug. 25,201 cases were processed. On Aug. 27,182 cases were on the docket. In August, the courtheard 104 drunken driving dispositions, 89 of which < val lug 6 lalanca ma Clink, UC Larry Taylor Debby Taylor National Board for Certif ication in Hearing Internment Science! 830-627-8660 ADVANCED HEARING CARE WITH THE ATTENTION YOU DESERVE Common Street Professional Bldg. • 1583 Common Street, Suite IOO • New Braunfels, Texas 781 30 MARK DuFRENE/KRT California Gov. Gray Davis and his wife, Sharon . wave to supporters after voting in West Hollywood. Calif.. Tuesday. ;
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The Secrets is a British drama series that focuses on the moral dilemmas that have to be resolved after secrets are unearthed over a series of stand alone dramas. Stars featured are Olivia Colman, Alison Steadman and Steve Oram.
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Microsoft Silverlight 3 launch event set
Microsoft is planning a launch event on July 10 for its Silverlight 3 technology and the accompanying Expression Studio 3 toolset, a company representative said Friday evening.
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Fire extinguished near Highway 118 and Rocky Peak Road
A small brush fire was put out in rugged terrain near Highway 118 and Rocky Peak Road in the Simi Valley area.
The fire was reported at about 6 p.m. and extinguished 25 minutes later, according to Erik Scott, spokesman for the Los Angeles Fire Department. Firefighters used water drops from a helicopter because of the fire's location, Scott said.
No structures were threatened and no injuries were reported, he said.
The Ventura County Fire Department and Los Angeles County Fire Department also responded to the fire, which covered about 100 square feet.
The cause of the fire was under investigation.


text:
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Feeling like a real geek
2 replies 361 views Edited Jun 18, 2012
I recently started to use the drake ring that came with my UC3 collectors edition and every time I walk outside and see a ledge I want to climb it just like drake. Do you guys have any video game accessories that you actual use outside of your house?
"I'm not a failure because I didn't win, I'm a failure because I didn't try." - Kev
Youtube: Akirasolid Entertainment
MVP Support
Registered: 05/12/2009
11041 posts
Re: Feeling like a real geek
Jun 22, 2012
Not me, i just collect them
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