<% function getFileName() Dim pos, path, name path = Request.ServerVariables("PATH_INFO") pos = InStrRev(path,"/",-1,1) name = Right(path, len(path) - pos) 'response.write("PATH_TRANSLATED" + Request.ServerVariables("PATH_TRANSLATED") + "
") '**** Test Code *********************** getFileName = name 'Response.Write("filename for NS6 " + name + "
") 'Response.Write(strMainPhysicalPath & Replace(Request.QueryString("NAME"),"%2E",".")) End Function ' Function IsEmailValid(strEmail) ' Action: checks if an email is correct. ' Parameter: strEmail - the Email address ' Returned value: on success it returns True, else False. Function IsEmailValid(strEmail) Dim strArray Dim strItem Dim i Dim c Dim blnIsItValid ' assume the email address is correct blnIsItValid = True ' split the email address in two parts: name@domain.ext strArray = Split(strEmail, "@") ' if there are more or less than two parts If UBound(strArray) <> 1 Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If ' check each part For Each strItem In strArray ' no part can be void If Len(strItem) <= 0 Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If ' check each character of the part ' only following "abcdefghijklmnopqrstuvwxyz_-." ' characters and the ten digits are allowed For i = 1 To Len(strItem) c = LCase(Mid(strItem, i, 1)) ' if there is an illegal character in the part If InStr("abcdefghijklmnopqrstuvwxyz_-.", c) <= 0 And Not IsNumeric(c) Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If Next ' the first and the last character in the part cannot be . (dot) If Left(strItem, 1) = "." Or Right(strItem, 1) = "." Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If Next ' the second part (domain.ext) must contain a . (dot) If InStr(strArray(1), ".") <= 0 Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If ' check the length oh the extension i = Len(strArray(1)) - InStrRev(strArray(1), ".") ' the length of the extension can be only 2, 3, or 4 ' to cover the new "info" extension If i <> 2 And i <> 3 And i <> 4 Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If ' after . (dot) cannot follow a . (dot) If InStr(strEmail, "..") > 0 Then blnIsItValid = False IsEmailValid = blnIsItValid Exit Function End If ' finally it's OK IsEmailValid = blnIsItValid End Function %> <% if LCase(getFilename)= "inthelab.asp" then response.write("Work being done in the Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI") end if if LCase(getFilename) = "labmembers.asp" then response.write("Staff, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI") end if if LCase(getFilename) = "publications.asp" then response.write("Publications, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI ") end if if LCase(getFilename) = "contactus.asp" then response.write("Geller Lab Contact, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI ") end if if LCase(getFilename) = "location.asp" then response.write("Nabel Lab Location") end if if LCase(getFilename) = "imagegallery.asp" then response.write("Geller Lab Images, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI ") end if if LCase(getFilename) = "favoritejournals.asp" then response.write("Favorite Journals, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI ") end if if LCase(getFilename) = "macroanalysis.asp" then response.write("Macro Analysis, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI ") end if if LCase(getFilename) = "labmethods.asp" then response.write("Lab Methods, Developmental Neurobiology Lab, Div. of Intramural Research, NHLBI ") end if %> - NIH <% Dim image_intheLab Dim image_labmembers Dim image_publications Dim image_contactus Dim image_location Dim image_imageGallery Dim image_nabelLabHOme Dim image_favoriteJournals Dim image_labMethods image_intheLab = "arrow_off.gif" image_labmembers = "arrow_off.gif" image_publications = "arrow_off.gif" image_contactus = "arrow_off.gif" image_location = "arrow_off.gif" image_imageGallery = "arrow_off.gif" image_nabelLabHome = "arrow_off.gif" image_favoriteJournals = "arrow_off.gif" image_labMethods = "arrow_off.gif" if LCase(getFilename)= "inthelab.asp" or LCase(getFilename)= "indexns6.asp" then image_intheLab="arrow_on.gif" end if if LCase(getFilename) = "labmembers.asp" then image_labmembers = "arrow_on.gif" end if if LCase(getFilename) = "publications.asp" then image_publications = "arrow_on.gif" end if if LCase(getFilename) = "contactus.asp" then image_contactus = "arrow_on.gif" end if if LCase(getFilename) = "location.asp" then image_location = "arrow_on.gif" end if if LCase(getFilename) = "imagegallery.asp" then image_imageGallery = "arrow_on.gif" end if if LCase(getFilename) = "nabelLabeHome.asp" then image_nabelLabeHome = "arrow_on.gif" end if if LCase(getFilename) = "favoritejournals.asp" then image_favoriteJournals = "arrow_on.gif" end if if LCase(getFilename) = "labmethods.asp" then image_labMethods = "arrow_on.gif" end if %>
Geller Lab Home Jump pass to content National Heart, Lung, and Blood Institute
Developmental Neurobiology 		Jump pass to links
[Border] <%if image_inthelab="arrow_on.gif" then%> In the Lab <%end if%> <%if image_labmembers="arrow_on.gif" then%> Lab Members <%end if%> <%if image_publications="arrow_on.gif" then%> Publications <%end if%> <%if image_contactus="arrow_on.gif" then%> Contact Us <%end if%> <%if image_location="arrow_on.gif" then%> Location <%end if%> <%if image_imageGallery="arrow_on.gif" then%> Image Gallery <%end if%> <%if image_nabelLabHome="arrow_on.gif" then%> Geller Lab Home <%end if%> <%if image_favoriteJournals="arrow_on.gif" then%> Favorite Journals <%end if%> <%if image_labMethods="arrow_on.gif" then%> Lab Methods <%end if%>  
Table image of part of Helix NHLBI Home NHLBI Labs    
arrow image to point to Nabel Lab splash page<%if image_nabelLabHome="arrow_off.gif" then %><%end if%>[Nabel Lab Home]<%if image_nabelLabHome="arrow_off.gif" then %><%end if%>arrow image to point to in the lab page<%if image_intheLab="arrow_off.gif" then %><%end if%>[In the Lab]<%if image_intheLab="arrow_off.gif" then%><%end if%>arrow image to point to in the lab members page<%if image_labmembers="arrow_off.gif" then %><%end if%>[Lab Members]<%if image_labmembers="arrow_off.gif" then %><%end if%>arrow image to point to publications page<%if image_publications="arrow_off.gif" then %><%end if%>[Publications]<%if image_publications="arrow_off.gif" then %><%end if%>arrow image to point to Contact page<%if image_contactus="arrow_off.gif" then %><%end if%>[Contact Us]<%if image_contactus="arrow_off.gif" then %><%end if%>arrow image to point to lab location page<%if image_location="arrow_off.gif" then %><%end if%>[Location]<%if image_location="arrow_off.gif" then %><%end if%>arrow image to point to Favorite Journals<%if image_favoriteJournals="arrow_off.gif" then %><%end if%>[Favorite Journals]<%if image_favoriteJournals="arrow_off.gif" then %><%end if%> arrow image to point to Lab Methods<%if image_labMethods="arrow_off.gif" then %><%end if%>[Lab Methods]<%if image_labMethods="arrow_off.gif" then %><%end if%>

 

Defrosting Eukaryotic Cells Frozen In Liquid N2

    Reagents / Solutions

    • Frozen cell line from liquid N2 tank..
    • Appropriate growth media (e.g., DMEM/10% FCS)
    • Fetal calf serum (FCS).

    Protocol  (6/1/00)

    1. Make 20ml growth media/10% FCS for each vial of cells to be thawed.
    2. Remove vial(s) of cells and thaw rapidly by hand suspension in the 37 degree water bath.
    3. Once thawed, add media/10% FCS  to the vial very slowly.  First ml should take ~ 1 min, second ml should take ~ 45 seconds etc. Triturate cells and then transfer to a 10 ml conical centrifuge tube.
    4. When cells are resuspended in 10ml spin down in bench-top centrifuge and resuspend in 10 ml media/10% FCS to get rid of remaining DMSO (note that this step has been omitted by some labs).
    5. Place cells in a T25 flask and allow to settle
    6. Allow to grow overnight.
    7. Assess cell number and viability, if the cell density is too high for the normal growth of the cell line split cells into multiple flasks.
Table image of part of HelixTable image of Helix

 

Dr. Herbert Geller, Head
Developmental Neurobiology, NHLBI, NIH
Building 10, Room 8C-213, 10 Center Drive, MSC-1650
Bethesda, MD 20892-1650
Email: gellerh@nhlbi.nih.gov
NHLBI HomeLabs at the NHLBIContact the NHLBIAbout NHLBISearch NHLBI
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