scholarly journals Methods development for measuring and classifying flammability/combustibility of refrigerants. Interim report, task 2 - test plan

1994 ◽  
Author(s):  
E.W. Heinonen ◽  
R.E. Tapscott
Keyword(s):  
Interim Report ◽  
Test Plan ◽  
Methods Development

En face dual fluorescence staining of fatty streaks allows observation of initiation and progression of atherosclerotic lesions

1995 ◽  
Vol 53 ◽  
pp. 864-865
Author(s):  
Anne M. Klinkner ◽  
Crystal R. Waites ◽  
Peter J. Bugelski ◽  
William D. Kerns
Keyword(s):  
Fluorescent Dyes ◽  
Nile Red ◽  
Dimethyl Formamide ◽  
High Cholesterol Diet ◽  
Methods Development ◽  
Atherosclerotic Lesions ◽  
En Face ◽  
Biochemical Evaluation ◽  
Stock Solutions ◽  
Dual Staining

A primary effort in the understanding of the progression of atherosclerotic disease has been methods development for visualization of the atherosclerotic plaque. We introduce a new method for the qualitative analysis of lipids in atherosclerotic fatty streaks which also retains those lipids for biochemical evaluation. An original aspect of the process is the ability to view an entire fatty streak en face, selectively stained for specific lipid classes within the lesion.New Zealand white rabbits were fed a high cholesterol diet(0.15%-0.3% for 14 wks). The aorta was removed and fixed in Carson's phosphate buffered formaldehyde followed by dual staining in the fluorescent dyes Nile red and filipin. Stock solutions of nile red(0.5mg/ml acetone) and filipin(2.5mg/ml dimethyl formamide) were prepared and kept at -20°C; all subsequent steps were at RT. 0.5cm × 1.0cm pieces of aorta were trimmed and adventitia removed. The pieces were then washed 3×15 min in PBS w/o CaMg, soaked in Nile red(NR)/filipin(Fl) stain(100(il NR stock + 200μl Fl stock in 10 ml PBS for 30 min, washed in PBS 3×30 min, rinsed with distilled water, mounted(Crystal Mount, Biomedia) and coverslipped and viewed by fluorescence microscopy.

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