[Histonet] Cryosectioning of retina/choroid/sclera?

[Srikant Tripathy]

Hi,

I am Srikant a PhD student in Queensland University of
Technology,Brisbane,Australia.In a part of my study I am trying to estimate
the element concentration (Na,K,Cl) at various layers of eye
(retina,choroid,sclera) by EDX analysis and confocal  microscope.

I am a Pharmacist and don't have very good knowledge of histological
techniques.

 

 I am using cryosectioning of eye layers and for a fragile layer like retina
I need to cryoprotect before sectioning. I am using snap freezing
technology.

 

My doubts are:

1.	As I can't use water soluble cryoprotectant as this will leach all
soluble ion. I wonder how people can keep retina soaked in Ames's solution
and gets fluorescence for chloride ions using dyes like MEQ. 
2.	Can any one please guide me how I will proceed for cryoprotecting
and sectioning the retina/choroid/sclera with out adverse effects on
elemental concentration? 
3.	Can I use PEG as a coating material prior to cryosectioning for
elemental analysis before EDX and confocal microscopy? 

 

Thanks.

Srikant

 

 

Srikant Tripathy | PhD Student | Institute of Health and Biomedical
Innovation 

Queensland University of Technology | Corner of Musk Avenue and Blamey
Street, Kelvin Grove QLD 4059 Australia

t: +61 (0) 7  3138 6157 | f: +61 (0) 7 3138 6030 | m: 0423586276 | e:
s.tripathy <@t> qut.edu.au <BLOCKED::mailto:s.tripathy <@t> qut.edu.au>   |  w:
<BLOCKED::http://www.ihbi.qut.edu.au/> www.ihbi.qut.edu.au

 

CRICOS No. 00213J