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ĤƬǯϵͳĵѧе𶯡ͻеƯƵķ봦

ʱ:2012-08-05 08:50Դ: :δ֪ Ķ:

ĤƬǯϵͳĵѧе𶯡ͻеƯƵķ봦

 

ժ Ҫ ĤƬǯͨ¼ϵͳǵоҪߣĿǰѾչΪӵĹӦϵͳӦùձŵѧ(ĤƬǯϵͳڵ)е(¼缫˵Ư)ϸ۲Щ⣬ЧĽϵͳŻ

ؼ: ĤƬǯѧе𶯣缫Ư

ĤƬǯͨ¼ϵͳоϸĤ(Ĥ)˫ӲǶͨͨ͸ԵҪ[1]ԽԽʵĤƬǯ¼豸㷺ϸĤͨIJϸڡҩѧѧ񾭿ѧԿѧϸѧѧȸ[5]

ĤƬǯϵͳѾӴͳĤƬǯѧϵͳչΪӵϼϵͳĤƬǯϸ¼ϵͳĤݼ¼ϵͳͷϵͳӫϵͳҩϵͳ΢ϵͳȶϵͳ[4]͸ŵĴҲøˡڹڵĤƬǯʵҽйװľĤƬǯϵͳձڵĵѧ(ĤƬǯϵͳڵֲ)е(¼缫˵Ư)ϸЧĴϣܶԹĤƬǯĿԱ.

1. ѧŷ

ѧɷΪŴڲԴԷŴԴԴһ㶼Ƿdz͵ģ¹HEKA˾EPC-9,EPC-10ָ꣺̽ͷգŴBessel˲ѡDC-3 kHz< 0.08 pA rms[8]Axon˾AxoPatch200Bʹ̽ͷ似ͬ£ɵʹ< 0.06 pA rms[6]

ǰĵѧɹΪԴĤƬǯϵͳҪٵǴӷŴ̽ͷ˿ʼ¼۲죬ɷŲϵ缫гϲ΢缫ٷõϷˮзӵȲĽУԴӡǿɰԴΪػ·ŸӦ[2]

1.1 䴦ʩ

һҪҪʵҸԴ͵ԴĹƵ(50Hz)ĸƵַ豸ֻƵȡͨӵصĵβϷ·ʹЧʵϵͳⲿ󲿷ֿռϸĴʩĺۡ

1.2 ػ·䴦ʩ

β赲̽豸β㱾ȴΪ˷˵ɣЩҪͨ“Ӵ”ͷšβγɹУʱҲ“ӵ”ʵʵĺǽβýγɵȵλβԽ͵γɵȻһβŴźŵ󽫷ŴźŵǽϽĵԴ(ȫ)򵥶ӵ[3]ҪֶӵأӵػڲͬĽӵصϸ΢ĵλӶβвγɵػ·һֻ·ųͿܽһǿ·

1.3 ŸӦ䴦ʩ

һ仯Ĵͨиһ·ȦʱͻᷢŸӦ·впڵԴĵĸͨ䲨ʶ䲨зҵ״ƵǹƵ(50Hz)ĸߴгԴԶе·ⲻܣʹ˫ͨиԽŴźŵ̽ͷ¹˫ߡ

2. ͵ѧIJ

2.1

HEKA-EPC9/10ĤƬǯ¼ϵͳн˲ԡȹһӵص“ȵλϵͳ”׼һͭӺͭ“λڵ”ȻNewport̨桢OlympusIX71΢塢Burleigh-PCS5200΢ٿ΢ٻܡMPSҩϵͳͼҩϵͳܵ豸“”ͨһ(ֻһ)ӵ“λڵ”ϡٽŴ“źŵ”ͨһӵ“ȵλڵ”ϡǶԷŸӦػ·˳ĴȻڸ¶ϵͳгԣͼ1-1ݼ1Fig.1-1СƫߡжϴΧʰȡҪԭڵĸ豸ǶķԴڵЧӦijЩλûַǿ㡣Ϊ˽ЩԵ缫λ(һϷ΢۹⾵·)Ӱ죬ǿchamber͵缫ΧٷõڶΡ

2.2 ǿֲ

һ赲chamber·΢ϵijЩչݿףӵصβ쵽chamber·ֻ㹻Ŀռƶchamberýӵֽλ̽ͷ(ͼ1-2)ĸƼ1Fig.1-2СڶΪchamberϷĸšýӵֽγ©״ֻһͨ(ͼ1-3)ĸƼ1Fig.1-3СǽchamberĴ󲿷λýӵصֽchamberΧ©״ֽͣγһĽӵش(ͼ1-4)ĸƼ1Fig.1-4УõԵĸơ


ͼ 1. ̽ͷԵŻ̡

1-1ʼ̽ͷλã1-2,1-3chamber·Ϸ1-4chamberȫΡ

1. ͬµ(λpA RMS)



2.3 λڵĸԴ

̽ͷתӵδıԵʱ뿪ֲεıǹ۲쵽ӵ1.2pA RMSڻڽǿķԴڵĸָ豸΢΢١ҩϵͳϵͳDZڵķԴЩ豸ӦʹýĻڲԪΣҲֱ硣ʹý豸ʽϴ豸Ӧܷõ棬򾡿̵ԴڵijȡȻ΢ԴֱԴ൱ɹ۵ĽϡԴĵǿȱ仯Ƶʱ仯ԵķֻҪܣӦʹϸֽȰԴߣγһӵβ㡣ҩϵͳעҺchamberĹҲҪΡ̽ͷŴ̽ͷ£ٽΣҽŴźŵ̽ͷ¹˫ʵ¡еź߶ӦʹôεBNC£β㶼뵼ͨγɵȵλ㲢ӵһӵضϡЩʩϿҪķš

ǽ̽ͷΪһ̽ߣڵֲͷ򣬷ֽӹϼϵͳеĹܷϴڽܵĿ͵Դֽ1.2pA RMS½0.57pA RMSƵķϵͳϸļͳΣ½0.215pA RMS̽ͷصֲαΧʱָ0.11pA RMSˮƽ

2.4

ֲʱҪעֶӵءʵʲУڲڲĽӵص·¶ӵءBNCβͨźŵػͨͬźŵؿܱڷĵԴϡϿBNCβ߶ϿijЩԴߣͿ·

ĤƬǯϵͳ“ӵض”Էֳ֣һźŵ(Signal GND)һɷŴṩǷŴźŵ0Vο㡣һֻ(Chassis GND)һ䵼ͨЩ豸ûԤúõĻأʱѡij¶˿ӵ㡣һо“Դȫӵ”ͨϵͳ“Դȫӵ”ǽӴصģҲͽӴˡǿҪ鹩ϵͳ“ȫ”ǷĽ˴ءΪǷЩʵҵ“Դȫӵ”յģǿ豸Ҳʹôյ“Դȫӵ”һǿ豸©磬ͻʹ“ȫ”豸в

3. еŷ

ĤƬǯϵͳУµ缫˵Ļе𶯺ƯҪĻеš

3.1 ϵͳ

𶯻ֱӵ·ʧܺͼ¼𶯻ζҪԵĴֱ𶯡񶯡ûܵķ̨ЧԵĴֱ[2]ڷ̨Ϸĸ𶯲ţڲʹһϩϻϿĩϣ񶯵Ӱ졣ʵڵңѡҹʵ飬ԱԴ

3.2 ֹϵͳƯ

ĤƬǯʵʹõļ¼΢ٱ̽գƶ(΢١г缫ˡchamberеϸ)Ӧ̡ܶ΢ӦֱӰװ΢ӿƽ̨ϡŴ̽ͷӦֱӹ̶΢ϣҪͷؽĽṹֹBurleigh˾PCS5000/6000ϵѹ羧΢ٺSutter˾MP285ϵ΢ܶdzҺѹ΢ҺϵԵʸ׷ƯơʵУ𶯸ײܼʱԻĵ缫˵Ưƣѵö࣬Ҫܳʱ۲ж[9]ƯƿɶΪ缫ϸƶڸ߷ŴᄉCCDϵͳпԹ۲쵽缫ƯƵԭǵ缫̽ͷг΢١̶װá΢塢΢ƽ̨ϸһ۲쵽ƯƣҪ۲жϣԶλƯԭ򡣸ǶPCS5000MP285΢ٵķʵʹþ飬һɰ˳飺

3.2.1 ¶ȱ仯ʹõIJϡ

¶ƯƵһҪԭ罫ƿ̽ͷгƵ¶ȾԲԵƯơҪʹϵͳܿյڡգԼκԴʹ¶ȱ仯С缫λйصĻеӷΧڱʹ¶вۺϲϡʵϵʯӢ0.56β3.21222ʯӢȶ[11]Ҳиȵʣܿɢȡʹͬһֲʱ¶ȱ仯ɵһµģڲԵƯơijЩе£Ҫ÷ʵԱ¶ϵͳӰ졣

3.2.2 еĻеӡ

ǿץס̽ͷ΢зؿƶָоϸ΢Ŀ˵ϵͳijɶһô΢۲缫ˣ缫ֲԹ̶΢ٵƽ̨

3.2.3 ̽ͷ͵缫г

ж̽ͷ缫ֱӹ̶΢ϣɼ缫ǷƯơǸݾжϣƯƶ̽ͷ͵缫гµġΪʩӸѹĽƤܶԼгӦҪѡ񾡿ļӽƤ̶ܲ̽ͷӰ졣Axon˾ڵ̽ͷHL-1/2ʹõƹ̶ļг̶׶ظ¡ͷͲĦڶʹúĦظ¡ζʹӱòȶAxonĽHL-UʹӣHL-Uʱмɹ̣ظ¡ƵĻɳڻᵼصƯơHEKA˾ļгdzȶHEKAʹһBNCͷ̶г̽ͷϡϽܣеȶԷdzáHEKAӳññвһԲԲԾȷ͹̶OȦڼгڵĵ缫ƶڽβ˵OȦԷֹгBNCƶƻ֤˵ʩӸѹʱ缫ƶ

3.2.4 µӦ

Ҫȷ΢ٸܹȫƽƶԼ΢Ƿƶû谭Ҫ֤еɷõģûи΢ʩ缫ʱ΢ٺ̽ͷ¶ŤʣӦЩӦͷʱͻᵼƯơ۲쵽Ưڼм΢ףƯƿܾԵӦǿɽ΢ٵ̳ĴȦʹе΢ֱ˶ת˶Χڶġ΢ʱ²ܱѹ졣ǻԽеĵ¶ֱ֧΢ٵϷСӦ

4. С

ҪĤƬǯ¼ϵͳеĵѧͻе⡣ϸ˸ָŵԴЧĽʩǽЩװԶHEKAAxonϵĤƬǯϵͳľܽᡣǵķۺʹڹĤƬǯʵҽһŻϵͳвοֵĤƬǯͨ¼Ե𶯺͵ƯƵҪǷdzߵģ˱Ķڳ¼ϵͳҲһο塣



[1] E Neher ,B Sakman. The Patch Clamp Technique. Sci . Amer . ,1992 ,Mar :44 51

[2] Rivka Sherman-Gold,, The Axon Guide for Electrophysiology Techniques, Axon Instruments, Inc.,1993,19-23.

[3] ר,.ĤƬǯϵͳ[J ] ; йҽѧѧ,1992 ,11(2) :128 - 137.

[4] .ϸĤƬǯ[J ]; йҽе־, 2000, 24 (3) : 155- 160.

[5] ¦ѩ,ר.ⵥͨȫϸ¼[J]; йҽе־, 2000, 24(4): 221- 226.

[6] Axon, Axopatch200B_manual_Rev_D, 2001, Specifications, 103-105.

[7] Axon, MultiClamp700B_manual_Rev_C, 2004, Specifications, 141-146.

[8] HEKA , Low-Noise Recording, Heka EPC10_manual, 2003, 66-68.

[9] Pipette Drift Sources and Solutions for Patch Clamp Recording Experiments, Burleigh Instruments , Inc. Application Note

[10] Richard A, Rae, JamesL, Low-Noise PatchClamp Techniques. Methods Enzymology. 1998, 293:218-266

[11] F.Sachs., A Low Drift Micromanipulator Holder, European Journal of Physiology 1995, 429:434-435.

 

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