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CLSM volume rendered images and media files of Malagasy Conostigmus.
This image set contains the volume rendered media files and images used in the manuscript: Mikó et al. (in press) Malagasy Conostigmus (Hymenoptera: Ceraphronoidea) and the secret of scutes. Peerj
Sample preparation
Male genitalia were temporarily mounted between two coverslips (1.5 µm, 22$\times$60) in a glycerin droplet, which did not reach the edge of the coverslip. We used Blu-tack (Bostik, Wauwatosa, WI, USA) as spacer as this material does not interact with glycerol and provides an adjustable, appropriate distance between the coverslips. Specimens were examined with an Olympus FV10i desktop CLSM using the a 60X objective.
Comments
Soft and sclerotized anatomical structures in arthropods tend to fluoresce with different intensities at different wavelength intervals \citep{miko_what_2013}. CLSM tissue-specific contrast is gained by exciting specimens using multiple excitation wavelengths and/or recording the fluorescence on multiple channels assigned to different laser wavelength intervals. In previous research, specimens were excited with only one blue laser (480 nm) and the auto-fluorescence was detected with two channels (500--580 nm and 580--800 nm). Although the resulting micrographs had differences in their intensity patterns, data from the two channels largely overlapped. In the present paper, we use two different lasers (631 nm and 499 nm) and set filters (644 nm and 520 nm respectively; narrow green and narrow red presets in Olympus Fluoview viewer software version 4.2) with narrow wavelength windows that result in a much higher tissue-specific contrast, almost perfectly separating muscle tissue and skeletal components.
Sample preparation
Male genitalia were temporarily mounted between two coverslips (1.5 µm, 22$\times$60) in a glycerin droplet, which did not reach the edge of the coverslip. We used Blu-tack (Bostik, Wauwatosa, WI, USA) as spacer as this material does not interact with glycerol and provides an adjustable, appropriate distance between the coverslips. Specimens were examined with an Olympus FV10i desktop CLSM using the a 60X objective.
Comments
Soft and sclerotized anatomical structures in arthropods tend to fluoresce with different intensities at different wavelength intervals \citep{miko_what_2013}. CLSM tissue-specific contrast is gained by exciting specimens using multiple excitation wavelengths and/or recording the fluorescence on multiple channels assigned to different laser wavelength intervals. In previous research, specimens were excited with only one blue laser (480 nm) and the auto-fluorescence was detected with two channels (500--580 nm and 580--800 nm). Although the resulting micrographs had differences in their intensity patterns, data from the two channels largely overlapped. In the present paper, we use two different lasers (631 nm and 499 nm) and set filters (644 nm and 520 nm respectively; narrow green and narrow red presets in Olympus Fluoview viewer software version 4.2) with narrow wavelength windows that result in a much higher tissue-specific contrast, almost perfectly separating muscle tissue and skeletal components.
