在 FCS Express 进行批处理时出现旧格式或无效类型库错误消息（如下图所示） Microsoft Office Excel 的结果是由于有关计算机上 Microsoft Office Excel 安装方式的问题而导致的。

问题是安装了 Microsoft Excel使用特定的语言，但 Windows 正在以不同的语言运行，这会导致其他程序尝试访问 Microsoft Excel 时出现问题。

为避免遇到此问题，请确保 Windows 设置为使用与安装 Microsoft Excel 所使用的语言相同。

您可以在 Microsoft 支持网站 https://support.microsoft.co 上找到此问题的摘要m/en-us/kb/320369.

FCS Express 提供三种批处理运行模式：

请参阅我们的批处理详细概述，了解有关说明和示例不同的运行模式。

发生这种情况是因为绘图的右边距截断了最右边 x 轴的一些文本。如果发生这种情况，您必须增加右边距和 x 轴之间的间距。为此，请右键单击该图并选择\"格式”。在\"格式”屏幕中，点击\"背景”类别。增加右边距的百分比。这将为您提供边距和 x 轴文本之间的更多空间。

显示带有流式细胞术数据的绘图时的一个常见问题是在轴标题中显示正确的文本。根据仪器以及采集过程中使用的设置，信息可能存储在 FCS 文件中参数的 Name 关键字或 Stain 关键字中。

在版本 3 中，可以单独更改每个图 - 在轴选项格式类别中，您可以从\"名称”更改为\"染色”，甚至\"名称 + 染色”或\"染色 + 名称”。

在 FCS Express 5 及更高版本中（以及在FCS Express 4)，您仍然可以进行该更改，但只需在插入绘图之前进行一次。您可以通过单击\"文件”选项卡和\"选项”命令来执行此操作。然后，点击\"数据加载”并选择\"FCS 文件选项”类别。将\"关键字”的值更改为\"用作参数名称”，然后单击\"确定”。

您可以阅读有关在我们的手册中设置轴标签：

注意：用户选项更改后，仅适用于新加载的 .fcs 文件数据或新的布局。要让现有绘图反映新的参数名称，请按照以下步骤操作：

新的布局参数名称首选项现已到位，无需再次进行更改。

包含更改轴的分步说明的教程标签和其他常用编辑的 FCS Express 用户选项也可用。

Reprintedfrom:

MoreprotocolsfromthelabP.J.Hansenhomepage

P.J.HansenandJ.G.BettsDairyScienceDept.,UniversityofFlorida,Gainesville32611-0701andDept.ofBIOLOGicalSciences,EastTexasStateUniversity,Commerce,Texas,75429

ThismethodwasoriginallydevelopedinthelaboratoriesofR.M.RobertsandF.W.Bazertoallowmetaboliclabellingofsecretoryproteinssynthesizeddenovobyculturedporcineendometrium(J.Reprod.Fertil.60:41-48,1980).Ithassincebeenusedtostudysecretionofproteinsandprostaglandinsbyendometriumfromthecow,ewe,mare,bitchandotherspecies.Thetechniqueisalsousefulforcultureofperi-implantationconceptusesandplacentaltissuesformetaboliclabellingstudiesandtoobtainconceptussecretoryproteinsforbiologicalstudies.

ThemediumusediscalledPigMEM,whichisamodifiedminimumessentialmediumsupplementedwithnon-essentialaminoacids,vitamins,insulinandadditionalglucose.Formetaboliclabellingstudies,specificaminoacidsareomittedorreducedinconcentrationtoincreaseincorporationofrADIolabelintonewly-synthesizedprotein.Cultureisperformedonarockingplatforminoxygen-richconditionstoincreasegaseousexchange.Forconceptuses,however,satisfactoryresultshavebeenobtainedwithanatmosphereof5%CO₂inairandwithoutuseofarockingplatform(J.Reprod.Immunol.18:271-291,1990).

PreparationofPigMEMThisrecipeisforpreparing10lofPigMEMusingastartingMEMformulation(cat.no.M7270fromSigma,St.Louis,MOorcat.no.410-2400fromGibco-BRL,GrandIsland,NY)thatisdeficientinleucine,lysine,methionineandNaHCO₃.Smallervolumescanbepreparedasneeded.

1.Inalargevessel,add9lofH₂0.DissolveenoughpowderedMEMtoprepare10l.Add30gglucose,15mgmethionine,52mgleucineand725mglysine-HClsothattherewillbe4,0.1,0.1and1.0timestheusualconcentrationsofthesesubstancesfoundinMEM.

2.Add22gNaHCO₃,100mlofGIBCOMEMvitaminsolution(cat.no.320-1120)orequivalent,100mlofGIBCOnon-essentialaminoacidssolution(cat.no.320-1140)orequivalentand2000IUinsulin.AdjustpHto7.1-7.3.Bringvolumeto10.0lwithwaterandfilter-sterilize(0.2µm)aliquotsof400mlintoautoclaved500-mlmediumbottles.StocksolutionsofPigMEMcanbestoredforatleast6moat4C.

4.Prepare100mleachof1.50g/lmethionine,5.20g/lleucine,and29.20g/lglutamine.Dissolveeachsupplementinwater,filter-sterilizeandstoreeachsupplementseparatelyat-20Cinsterile12X75tubes.Similarly,prepare4.0mlaliquotsofGibcoantibiotic-antimycoticsolution(ABAM;cat.no.600-5240)orequivalent.

5.BeforeuseofPigMEM,supplementsareaddedtopreparethemediumforspecificuses.CompletePigMEM,whichisusedforcultureswhenmetaboliclabellingwithaminoacidsisnottobeperformed,ispreparedbyaddingonealiquotofleucineandmethioninetoa400-mlbottleofPigMEM.A4mlaliquotofABAMisalsoroutinelyadded;thiscanbeomittedifthetissueofinterestissusceptIBLetoantibioticcytotoxicity.Porcineandovineendometriumhavebeensuccessfullyculturedwithupto8mlofABAM/400mlPigMEM.Forlabellingwith[³H]leucineor[³⁵S]methionine,omitthecorrespondingaminoacidsupplement.Mediumcanbestoredforatleasttwomonthsafteradditionofsupplements.Analiquotofglutamine(4ml/400mlmedium)shouldbeaddedbeforecultureifgreaterthanthreetofourweekshaveelapsedsincethemediumwaspreparedorglutaminewaslastadded.Mediumshouldbewarmedto~37Cimmediatelybeforeuse.

PreparationofTissueExplantsofendometrium.Removereproductivetractfromanimalatsurgeryorslaughter.Underalaminarflowhood,useasterilescissorstoexposetheendometrium.Rememberthattheoutsideoftractisnotsterile.Useaforcepstoliftuterineendometriumandascissorstoremovetheendometriumfromunderlyingmyometrium(becarefultodistinguishbetweencaruncularandintercaruncularendometriumforcowsandewes,whenpossible).Duringcollection,placestripsofendometriumintoa100mmpetridishcontainingPigMEM.Usingascalpelandforceps,cutendometriumintoexplantsofabout2-3mm³.Next,washexplantstwotothreetimesbyaspiratingoldmedium,addingfreshmediumandswirlinggently.Thisisacriticalsteptoreducecontaminationofmediumwithserumproteinsthatcaninterferewithtreatmentsandanalysisofconditionedmedium.Formaximumreductioninserumproteincontamination,incubateexplantsinmediumfor2h,replacewithfreshmediumandthenaddradiolabelandtreatments.

Topreparecultures,addrequiredamountofmediumtoacleanpetridish(15mlina100x15mmPetridishfor500mgculturesor7.5mlina60x15mmPetridishfor250mgcultures).Placethedishonabalance,tare,andaddtissueexplantsuntildesiredweightisobtained.

Peri-implantationconceptuses.ThesecanbeobtainedbyflushingtheuterususingatranscervicalFoleycatheter(cattle),atlaparoscopy,orafterremovaloftheuterusatslaughterorhysterectomy(seemethodinthisvolume).TheembryoshouldbewashedwithPigMEMasdescribedforendometriumandculturedwhole(15mlmedium)orinhalves(7.5mlmedium).

CultureProcedures1.Ifrequired,add50-200µCiradiolabeledcompoundperdish.Isotopesshouldbeofthehighestspecificactivityavailableanddissolvedinaminimumamountofethanol.IsotopesthatcanbeusedincludeL-[4,5-³H]leucine(inleu-deficientmedium),L-[³⁵S]methionine(inmet-deficientmedium),D-[⁶H]glucosamineandD-[³H]mannose(forlabelingglycoproteins),L-[1-¹⁴C]leucine(forusewith³H-labelledsugarsfordual-labelling)andH₃³²PO₄(inaPO₄-freemediumsuchasSigmacat.no.M4898).SeeBiol.Reprod.36:405-418(1987)andBiochim.Biophys.Acta966:56-64(1988)forexamples.

2.Culturesareperformedat37ConarockingplatformsuchasthatsoldbyBellco(Vineland,NJ)inanatmosphereof50:45:5N₂:O₂:CO₂(v/v/v).Thisgascanbespecial-orderedorpreparedbymixingequalvolumesof100%N₂anda90:10O₂:CO₂mixture.Culturescanbeperformedingasboxes(cat.no.7741-10010,Bellco)toreducegasusage.Placethedishesintheunsealedbox,allowgastoflowintotheboxfor15min,sealtheboxandplaceintheincubatorontherockingplatform.3.Cultures,usuallyterminatedafter24hbycentrifugation,cancontinuefor3-4difmediumischangeddaily.Productsinthetissueandmediumcanbeenanalysedbyelectrophoresisandfluorography,immunochemicaltechniques,HPLC,bioassay,etc.