[histonet] inclusion of lipids drops in Imunoluorecsece of frozen section

[janick.jean-marie]

 Hi fellows  !!!

I have a prolem with an immunofluorescence on frozen section of mammary gland ... 
I can see lipid inclusion (lipids drops)... in my section when i look my immunofluorescence
I use this protocol ;

1.  Formalin 4% for 15 min
2. Bath of PBS 1x  for 15 min 

3. Antiboby A? against Beta-GAl(1/500) and against SCA-1(1/300)
4. 1 hour in humid chamber 
5. Bath of PBS 1x  for 15 min
6. 2 flurorescent secondary antibody (1/300)  (Rhodamin and Fluorescein)
7. humid chamber for 30 min 
8. wash with PBS 
9.  add mowiol for mountig scale
10. Stock at  -80°C

Have you got an idea for explain my problem ... and what can i do for remove the lipids drops ? 


 THANKS !!!
Janick JEAN-MARIE
CRLC INSERM 

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