Page 335 - Fundamentals of Light Microscopy and Electronic Imaging

318 APPENDIX I

magnified virtual image because the linear extensions of rays, focused on the retina by
the ocular and eye lens, appear to converge in a virtual image plane located about 25 cm
in front of the eye. The ray diagram in Figure 1-2 helps clarify the role of perspective in
perceiving a virtual image.

Chapter 6

(2) For a 40 0.65 NA dry lens and 546 nm green light, striae or pore spacings of 0.5
m are barely resolved. Only Gyrosigma, Navicula, and Stauroneis are clearly resolved.
Nitzschia and Pleurosigma are barely resolved. (3) The spatial resolution is given as d
0.61λ/NA (0.61 0.546)/0.65 0.51 m. (5) The diffraction patterns preserve
the orthogonal and hexagonal arrangements of striae and pores seen in the normal
(orthoscopic) viewing mode of the microscope. The narrowest spacings in the object
correspond to the widest spacings in the diffraction patterns. (6) Lowest resolution with
red light and restricted aperture; best resolution with blue light and open aperture. For
demonstration of the effect of condenser aperture size on resolution, try Gyrosigma,
Navicula, or Stauroneis with a 20 lens in green light; alternatively, examine
Gyrosigma, Stauroneis, Nitzschia, or Pleurosigma with a 40 lens in red light. (7) If a
100 oil lens is used without oil, the NA corresponds to about 1, and Pleurosigma is
barely resolved. With oil immersion, definition and resolution are both dramatically
improved and pores are evident.

Chapter 7

(3) The molarity of hemoglobin is calculated: g/L BSA at 50% match point 1/64,000
g/mol mol/L. The concentration of hemoglobin in erythrocytes is approximately
4–5 mM. This is 2,500 times the molarity of tubulin and 250,000 times the concentra-
tion of aldolase. (4) The range of phase densities is partly due to real differences in
hemoglobin content. The ratio of blood volume to BSA solution volume should be very
small and the solution should be mixed very well. Cells could be fractionated on a den-
sity gradient and the hemoglobin content of cells in various fractions determined quan-
titatively.

Chapter 9

(3) For the slow axis of the waveplate running NW–SE between crossed polars and a
magenta-colored background: starch gains, NE–SW quadrants yellow, SE–NW quad-
rants blue; bordered pits, NE–SW quadrants blue, SE–NW quadrants yellow. (4) Carbon
bond alignments in amylose chains are arranged radially as in a pincushion; the carbon
bond alignments in lignin molecules in wood pits are arranged tangentially, like the lay-
ers of an onion.