Squash preparations and living cells showing chromosome arrangement in mitosis, meiosis I, and meiosis II in green lacewing (Chrysoperla rufilabris) males

In previously-studied species of insects in the order Neuroptera, sex chromosomes in males exhibit a phenomenon called "distance segregation," where the X and Y chromosomes are separated and associated with opposite poles from prometaphase I-anaphase I. This study aimed to determine the chromosome number and sex determination of a previously unstudied Neuropteran insect, the green lacewing Chrysoperla rufilabris (Burmeister, 1839).  Following determination of chromosome number and sex determination we examined images of fixed and living specimens to determine if distance segregation was observed. This dataset includes all of the raw images of living and aceto-orcein squashed cells that were observed to determine chromosome number, sex determination, and existence of distance segregation.

Methods
Testes of Chrysoperla rufilabris were removed from adult male abdomens.  

For aceto-orcein squashes, isolated testes were fixed in 3:1 ethanol/acetic acid for 5 minutes, rinsed 30 seconds in distilled water, and stained in filtered aceto-orcein (2.5 g orcein in 100 mL 45% (v/v) acetic acid) for 3 minutes.  Stained testes were placed on a slide in a drop of 45% acetic acid.  A coverslip was placed over the testes and uniform pressure was applied by hand on the coverslip. 

For living-cell preparations, testes were placed under a thin layer of Kel-F #10 oil (Ohio Valley Specialty Company, Marietta, Ohio) in a culture chamber on a coverslip.  Testes contents were spread under the oil and living cells were immediately imaged.

Imaging was done on a Nikon Eclipse TS100 microscope (Nikon Instruments, Tokyo, Japan) equipped with a 100X, 1.25 NA phase-contrast, oil immersion objective.  Images were collected either with a View 4K HD camera (Microscope Central, Feasterville, PA, USA), using InFocus software version x64 (Microscope Central), or using a Spot camera with the Spot 3.5.7 software (SPOT Imaging, Sterling Heights, MI, USA).