INJECTIONANDHOLDINGPipetteS

Theglasscapillarytubingusedshouldbethinwalled,borosilicateglasswithoutafibre.

e.g.ClarkElectromedicalInstruments

GC100T-15(=15cm)

GC100T-10(=10cm)

OUTSIDEDIAMETER:1.0mm

INSIDEDIAMETER:0.78mm

LENGTH:150mm

AustralianAgents:

SDRClinicalTechnology,213EasternValleyWay,MiddleCoveNSW2068(02)9582688

Glassshouldbeacidwashed:-

1.Placeglassina100mlmeasuringcylinder,containing1:1v/vSulphuricacid,Nitricacid,soakovernight.Thisisdoneinfumehood,makesureglovesandgogglesareworn.

2.Pouracidintoanothercylinder,thiscanbereused.

3.Washglassatleast10xwithMilliQH20then3xwithTravenolH20.Drainwell.

4.CovercylinderwithParafilm,pokeholesinitusinganeedleandplaceglassin60¡Covenuntildry.Thistakesafewdays.

Pipettepulling:-

FlammingBrownmicropipettepullerModelP-87

SutterInstrumentCo.USA

UsingProgram6(VH)

Heat=915Pull=20Vel=90Time=120Pressure=500

Injectionpipettes:-

Injectionpipettesshouldhaveaninternaldiameteronlymarginallylargerthanthestemcells(18-20um)andasharppointwhichallowstheneedletopenetratecleanlyintheblastocoelcavity.

Theglassiscutbyhandunderastereodissectingmicroscope,atamagnificationofX50,usingasharpscalpelbladeandalargeglasspetridishfilledwithsilicon.(Dow-corningSilgard).Theglasswillusuallybreakwithasharpbevelledendwhentouchedwiththeedgeofthescalpelblade.Theresultantpipetteshouldhavearelativelylongsectionattheappropriatedimater(18-20um).

Holdingpipettes

Holdingpipettesarebluntflame-polishedpipettesthroughwhichsuctionisappliedtoimmobilizetheblastocyst.Thesamepieceofglassiscut,onlyfurtheruptheshaft~200um.Thisiscutbyholdingtheshaftofglassonyourforefingerwithyourthumbandscoredwithadiamondpencilthenbrokenbyflickingthesurplusoffthetop.Acleanneatbreakisessentialotherwisetheblastocystdoesnotsitproperlyontheendandcomesoffduringtheinjectionprocess.

Theendofthepipetteisthenflamepolishedbyholdingitinamicroburnerflameuntiltheinternaldiameterisreducedto20um.(1-2divisions).

================  蚂蚁淘在线  ================

免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容

版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。