Images were captured on the Deltavision IX70 system (Applied Precision; Olympus) with a 100× objective lens (NA 1.40) and a camera (CoolSNAP HQ2; Photometrics) using softWoRx software at RT. 7 consecutive z-slices with an interval of 0.2 µm were acquired, then processed by deconvolution using the constrained iterative deconvolution algorithm within softWoRx. The deconvolved z-slices were projected using a maximum-intensity algorithm to form the final processed image. Meiotic chromosomes were surface spread as described previously (Dresser and Giroux, 1988 (link)), except that glass slides were not precoated with plastic. In brief, meiotic cells from appropriate time points were digested with zymolyase in the presence of 1 M sorbitol to form spheroplasts. Cells were gently washed once, and chromosome spreads were prepared by resuspending spheroplasts in buffered solution without sorbitol (22 mM MES) and 3% PFA. Cell suspension was spotted onto clean glass slides and left for 20 min for spread chromosomes to settle. The slides with chromosome spreads were washed with PBS once and subjected to standard immunostaining protocols. Primary antibodies used were Zip1 and Red1 (rabbit/mouse, 400-fold dilution, S. Roeder (Yale University, New Haven, CT); Sym et al., 1993 (link); Smith and Roeder, 1997 (link)). Secondary antibodies were Alexa Fluor 488 or 594 (Invitrogen).