A sample of men and women aged 75+ years (n = 27) was recruited and observed in a home-like environment.
The subjects were reared in isolation and observed in a dyadic test (10 min) at the ages of 28 30, 37, 42 45, 72 and 180 280 days.
Postelectrophoresis, gel was stained with Coomassie Brilliant Blue dye and observed in a gel-imaging system (Chromous Biotech, Bangalore, India).
The slides were stained with Picrosirius Red Stain (Polysciences, Inc., Warrington, USA) and observed in a normal transmitted light microscope and a polarized light microscope (Nikon, Tokyo, Japan).
On Christmas Eve , 1968 Apollo command module pilot Jim Lovell compared space to earth and observed in a famous broadcast from space that: "The vast loneliness is awe-inspiring and it makes you realize just what you have back there on Earth".
The cells were then washed 3 times in PBS and observed in a Zeiss epifluorescence microscope (Axioplan 2).
After gold fixation with 2.5% glutaraldehyde, samples were counter stained and observed in a Jeol electron microscope.
Samples were dehydrated to 70% (v/v) ethanol and observed in a stereoscopic microscope (Leica MZ16) and an Eclipse 600 microscope (Nikon) equipped with Nomarski interference optics.
Ultrathin sections were contrasted in uranyl acetate and lead citrate and observed in a Philips CM100 electron microscope.
Finally, the sections were dehydrated in ethanol, cleared in xylene, mounted and observed in a light microscope.
After washing in PBS, the sections were counterstained with DAPI, mounted in Mowiol and observed in a confocal microscope.
