NANOPHYSIOLOGY OF CELLS, CHANNELS AND NUCLEAR PORES - Life at the Nanoscale: Atomic Force Microscopy of Live Cells - page 122

(b)

(a)

(d)

(c)

Figure 6.4. Series of AFM images showing inside-out oriented isolated RBC
membranes. The size of scanned areas is (a) 80

s

80 μm, (b) 40

s

40 μm, (c) 10

s

10

μm and (d) 5 5 μm. Images are colour-coded, poly-L-lysine-coated glass is shown

in “dark blue” and the lipid bilayer membrane with proteins is shown in “light blue”.
Overlapping membranes are shown in “turquoise”, and multiple overlaps are shown
in “brown“.

s

6.1.5 Qdot-Labelled Anbodies

Monoclonal antibody against the C-terminus of CFTR in combination with
secondary Qdot-labelled antibodies were used for immunostaining. 10 Because
of their properties as particles with deined size and superior luorescence,
the Qdots were used as excellent AFM and luorescence markers for CFTR
localization. Fluorescence microscopy of the site-speciic Qdot-labelled
CFTR showed clearly the presence of CFTR on human RBC membranes
( Fig. 6.5a,c ) . The weak non-speciic autoluorescence of the membranes,
due to glutaraldehyde ixation, enabled us to visualize each RBC membrane.
However, the membranes from CF patients ( Fig. 6.5b,d ) showed drastically