For morphology imaging, 20 ml of LB agar was added per plate, and the plates were dried at room temperature for one day before use. For VpsO complementation, the agar contained 1 mM IPTG, for all other strains LB agar only was used. For colony morphology, the overnight cultures were diluted to 10−8 and grown at 25°C for 5 days. Colony morphology was imaged with the Zeiss Stemi 2000-C microscope equipped with Zeiss AxioCam ERc 5 s Microscope Camera. Morphology experiments were carried out with minimum two biological replicates.
Axiocam erc 5s microscope camera
Manufactured by Zeiss
Sourced in Germany
The Zeiss Axiocam ERc 5s is a microscope camera designed for digital image capture. It features a 5-megapixel CMOS sensor and supports high-resolution imaging. The camera is capable of live image display and can be connected to a computer for data transfer and analysis.
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Lab products found in correlation
5 protocols using axiocam erc 5s microscope camera
1
Imaging Bacterial Colony Morphology
2
Morphological Characterization of Bacterial Cultures
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For morphology imaging, 20 mL of LB agar was added per plate, and the plates were dried at room temperature for 48 h before use. The overnight cultures were diluted 1:200 for spot morphology analysis, and 3 µL was spotted on a plate with three spots per plate total. The overnight cultures were diluted to 10−9 for colony morphology, and spots and colonies were grown at 30°C for 72 h. For pellicle analysis, the overnight cultures were diluted 1:200, and 200 µL of resulting dilutions was placed in a clear 96-well plate (Thermo Fisher Scientific) and grown statically at 30°C for 24 h. Spot, colony, and pellicle morphology were imaged with the Zeiss Stemi 2000-C microscope equipped with Zeiss AxioCam ERc 5 s Microscope Camera. Morphology experiments were carried out with a minimum of two biological replicates.
3
Quantifying Fern Gametophyte Growth Dynamics
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Gametophyte area was quantified on day 0 and day 8 of each treatment by processing photographic data obtained from an Olympus CKX41 inverted microscope (Olympus Co., Tokyo, Japan) with a Zeiss Axiocam ERC5s microscope camera (Zeiss, Jena, Germany), using ImageJ software [43 (link)]. Ten fields of view (100× magnification) per replicate were randomly photographed, and the area of all gametophytes (female and male) present in each field of view was measured. The gametophyte average area was determined per replicate. If a female gametophyte became fertile and formed oogonia, the area of the oogonia was included. In contrast, if eggs and developing sporophytes were detected they were not included in the area measurements.
Relative growth rates (RGR) were estimated using the following formula:
Relative growth rates (RGR) were estimated using the following formula:
4
Trap Formation in Arthrobotrys oligospora
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A. oligospora only develops traps in low nutrient environments in the presence of nematodes (4). Consequently, A. oligospora strains were first cultured for 2 days in 3.5 cm diameter LNM-containing Petri-dishes. Then, 30 adult living C. elegans nematodes were placed on the Petri-dishes for 6 hour and subsequently washed out from the plates with M9 buffer (22 mM KH2PO4, 42 mM Na2 HPO4, 86 mM NaCl). After an additional 18 hours of growth, A. oligospora strains were imaged using a Zeiss Stemi 305 Stereo Microscope (Zeiss, Göttingen, Germany) and a Zeiss Axiocam ERc 5s Microscope Camera (Zeiss, Göttingen, Germany). Three 40X images of the mycelium (size 2.5 x 2.1 mm) per plate were captured at random for 6 plates per strain, yielding a total of 18 images per strain.
5
Fungal Conidia Isolation and Imaging
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Spore suspension of fungal strains was inoculated on 5 cm PDA (YPD for N. crassa) plates and incubated for 5 days at 25°C in the dark. Next, 2 ml of ddH2O was added to each plate and conidia were carefully scratched off from the surface of the mycelium. The ddH2O from the plate was first filtered through two layers of non-woven cloth (to exclude extraneous hyphae) and then transferred to a 2 ml centrifuge tube for centrifugation at 13,000 x g for 1 minute. For each tube, 1 ml of supernatant was extracted and 10 x 5 μl droplets of the remaining solution were placed on an LNM Petri-dish. Each droplet was imaged using a Zeiss Stemi 305 Stereo Microscope (Zeiss, Göttingen, Germany) and a Zeiss Axiocam ERc 5s Microscope Camera (Zeiss, Göttingen, Germany) at a resolution of 80X and 16X, for the conidiation study of fungal species and A. oligospora strains, respectively. The images comprised 2290×1920 pixels, representing an area of about 1.3 x 1 mm for the 80X images and 6.3 x 5.3 mm for the 16X images.
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