Axio imager m2m microscope

Cross sections were prepared by embedding each sample within a double layer of methyl methacrylate resin (Technovit® 2000 LC). Each layer of resin was cured under UV light for 20 min. Excess resin was ground off and the surface was finely polished using CarbiMet 2 and Micro-mesh abrasive paper of various grits to expose the sample’s stratigraphy. Cross sections were examined by means of a Zeiss Axio Imager M2m microscope, equipped with an Axiocam HRc digital camera and providing 50×, 100×, 200×, 400×, and 500× magnification. Photographs were collected using the AxioVision 4.X.X software.

Epifluorescent images were taken using a Zeiss AxioImager M2m microscope equipped with a Zeiss AxioCam MRm camera and AxioVision software 4.8.

Animals carrying the unc-25p::GFP reporter strain to visualize VD and DD motor neurons were synchronized and allowed to develop at 20°C and FUdR was added at L4 stage. At L4, D2, and D9 of adulthood, animals were imaged in a Zeiss Axio Imager M2m microscope and the number of animals showing axonal breaks in the VD and DD motor neurons were scored. Axonal breaks were defined as an area of discontinued fluorescence in either the ventral or dorsal nerve cords.

FESEM imaging is a swift and robust technique for morphological studies. This technique has widely been used for the identification of clay minerals⁵⁶ . Images are produced by the interaction of the samples with primary electron beams that identify any microscopic objects (up to nanometer scale) such as clay minerals, grain shapes, wall structures of organisms, or for topographic characterization of any micro size particles⁵⁶ . For this study, FESEM analysis was conducted along with the XRD analysis for the identification of attached clay minerals on the outer part of the shells⁵⁶ . Clay materials were separated from outer shells during ultrasonic cleaning. The sample preparation technique involved various steps like sample stubbing, mounting and imaging. FESEM imaging of the uncoated clay mineral sample was conducted by ZEISS SUPRA 55 under laboratory conditions. We used the published catalog⁵⁶ for clay mineral identification. Microstructure studies of the different fossil specimens were done in the same FESEM. Light microscopic observation was performed under the Zeiss Axio Imager M2m microscope.

GFP::LGG-1/Atg8 foci formation was visualized as described in Kumsta et al., 2017 (link). Briefly, L4 stage and day 1 adult animals were raised on HT115 bacteria at 20°C and imaged using a Zeiss Axio Imager M2m microscope with AxioVision v4.8.2.0 software at ×63 magnification. Two independent trials where at least 20 seam cells from 15 to 20 different animals were scored for GFP::LGG-1 puncta accumulation. In all cases scoring of puncta was blinded with respect to genotype.

Images of tricaine-anesthetized larvae at 3 dpf were captured using the Vertebrate Automated Screening Technology (VAST) Bioimager (Union Biometrica) mounted to an AXIO Imager.M2m microscope (Zeiss) with a 10x objective lens. Larvae were passed sequentially through a 600 μm capillary on the detection platform. Each larva was detected by software on the computer screen and oriented automatically for lateral and ventral side images with a pre-provided template setting in the software. VAST software (version 1.2.6.7) operated in automatic imaging mode with a 70% minimum similarity threshold, as described^{73 (link)}. Bright field lateral images were captured with the VAST onboard camera and a fluorescent signal from ventrally positioned larvae with an Axiocam 503 monochrome camera (Zeiss) and ZenPro software (Zeiss).