Three central spikelets per replication were collected from all 200 spring wheat genotypes during the third weeks of February 2021 and February 2022. The maximum temperature ranges observed during the third weeks of February 2021 and February 2022 were 23–26°C and 20–23°C, respectively (https://www.accuweather.com ). Anthers were removed with a sharp needle and stored at 4°C in a refrigerator for microscopic study of pollen viability. Modified ALEXANDER test (Dafni and Firmage, 2000 (link)) was used for pollen viability test. The stained pollen grains were recorded under a compound microscope (Olympus). The darkly stained pollens were fertile and lightly stained pollens were sterile. Pollen viability was counted as the ratio of number of stained pollen to total number of pollen grains and quantified as percentage (Prasad et al., 2006 (link)).
Compound microscope
Manufactured by Olympus
Sourced in Japan, Germany
The Olympus compound microscope is an optical instrument designed to magnify and observe small objects, such as biological specimens, in great detail. It uses a combination of lenses to provide high-resolution images of the sample under examination. The core function of the compound microscope is to enable detailed observation and analysis of microscopic structures and features.
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Lab products found in correlation
Htm ivos 2, by Hamilton Thorne (1 mentions)
Colony counter, by HiMedia (1 mentions)
Bh 2 stereoscopic microscope, by Olympus (1 mentions)
Bcip nbt, by Merck Group (1 mentions)
T7 rna polymerase, by Roche (1 mentions)
Anti dig antibody, by Roche (1 mentions)
Cue 2 image analysis system, by Olympus (1 mentions)
Uv cabinet, by CAMAG (1 mentions)
Silica gel type g, by Merck Group (1 mentions)
Automatic tlc coater, by CAMAG (1 mentions)
Tlc jar, by CAMAG (1 mentions)
22 protocols using compound microscope
1
Evaluating Wheat Pollen Viability under Climate Variability
2
Assessing Worm Growth under Fatty Acid Supplementation
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Synchronized L1 worms were pipetted onto the bacterial lawns. Body lengths were measured on day 4.
Liquid culture (fatty acid supplementation): Synchronized L1 worms were pipetted into each well, which contained S-basal media and a supplemental fatty acid. Body length was measured on days 4 and 6, by placing worms in an agarose padded glass slide with coverslip. To assess growth of larvae after internal hatching, a few adult worms were placed inside the well and the lengths of the progeny were measured after 10 days. To measure their lengths, the worms were placed on an agarose padded glass slide and covered with a coverslip. They were then imaged using an Olympus compound microscope with DIC filters.
Liquid culture (fatty acid supplementation): Synchronized L1 worms were pipetted into each well, which contained S-basal media and a supplemental fatty acid. Body length was measured on days 4 and 6, by placing worms in an agarose padded glass slide with coverslip. To assess growth of larvae after internal hatching, a few adult worms were placed inside the well and the lengths of the progeny were measured after 10 days. To measure their lengths, the worms were placed on an agarose padded glass slide and covered with a coverslip. They were then imaged using an Olympus compound microscope with DIC filters.
3
Morphological Analysis of Yeast Isolates
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In the present study, the morphology of cells of the selected isolates and their appearance on yeast extract peptone dextrose (YPD) agar media was examined [15–19 ]. The medium was autoclaved at 121 °C and 15 psi and poured on a petri dish and cooled. After cooling, the plates were inoculated with 48-hour-old yeast strain and incubated at 30 °C for 48 h. The following features of the appearance of cultures were recorded: texture, color, and surface of colonies.
Selected isolates Pa and Or were cultured in YPD liquid medium. The medium was autoclaved at 121 °C and 15 psi and cooled. Fifteen milliliters of the medium was distributed into McCartney tubes, then inoculated with a half loop-full of 48-hour-old selected yeast strain, and incubated at 30 °C for 3 days. The culture was examined for the growth visually on the surface of YPD liquid medium, and the shape of cells, by compound microscope (Olympus, Japan) [20 (link)].
Selected isolates Pa and Or were cultured in YPD liquid medium. The medium was autoclaved at 121 °C and 15 psi and cooled. Fifteen milliliters of the medium was distributed into McCartney tubes, then inoculated with a half loop-full of 48-hour-old selected yeast strain, and incubated at 30 °C for 3 days. The culture was examined for the growth visually on the surface of YPD liquid medium, and the shape of cells, by compound microscope (Olympus, Japan) [20 (link)].
4
Comprehensive Sperm Quality Analysis
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The post-thaw samples were immediately assessed for sperm motility and kinematic parameters using a computer-assisted semen analyzer (CASA; HTM IVOS II, Hamilton Thorne Biosciences, Beverly, MA, USA). Progressive motility (%), total motility (%), average path velocity (VAP, µm/sec), straight line velocity (VSL, µm/sec), curvilinear velocity (VCL, µm/sec), amplitude of lateral head displacement (ALH, µm), beat-cross frequency (Hz), straightness (STR, [VSL/VAP]×100), and linearity (LIN, [VSL/VCL]×100) were evaluated.
The morphology of sperm was assessed by staining with Diff-Quick (Arnaparn, Nonthaburi, Thailand) and analyzed by an HTM IVOS II CASA equipped with a Dimensions II Strict Morphology software system using Kruger’s strict criteria. A total of 200 spermatozoa were analyzed in each slide at ×400 magnification.
Sperm viability was assessed using 0.5% (w/v) eosin-Y dissolved in 0.9% NaCl. A 10-µL sperm suspension was mixed with 10 µL of 0.5% eosin-Y. Then, the mixture was placed on a glass slide and covered with a coverslip. The samples were immediately assessed for sperm viability using a compound microscope (Olympus, Tokyo, Japan). A total of 200 spermatozoa were analyzed in each slide [22 ]. The spermatozoa were classified as live (unstained heads) or dead sperm (stained red or dark pink heads) and reported as the percentage of live sperm.
The morphology of sperm was assessed by staining with Diff-Quick (Arnaparn, Nonthaburi, Thailand) and analyzed by an HTM IVOS II CASA equipped with a Dimensions II Strict Morphology software system using Kruger’s strict criteria. A total of 200 spermatozoa were analyzed in each slide at ×400 magnification.
Sperm viability was assessed using 0.5% (w/v) eosin-Y dissolved in 0.9% NaCl. A 10-µL sperm suspension was mixed with 10 µL of 0.5% eosin-Y. Then, the mixture was placed on a glass slide and covered with a coverslip. The samples were immediately assessed for sperm viability using a compound microscope (Olympus, Tokyo, Japan). A total of 200 spermatozoa were analyzed in each slide [22 ]. The spermatozoa were classified as live (unstained heads) or dead sperm (stained red or dark pink heads) and reported as the percentage of live sperm.
5
Microbial Cultivation and Analysis
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The solvents, sugars, staining dyes and other chemicals were used of analytical grade and purchased from either HiMedia or SD-fine Chem. Ltd. Mumbai. The instruments viz. colony counter (HiMedia, Mumbai), compound microscope (Olympus Corporation, Japan), cooling centrifuge machine (Remi, Mumbai), bacteriological incubator (Kumar Corporation, Mumbai), orbital shaking incubator (CIS 24BL, Remi, Mumbai), digital pH meter and thermometer (Systronics), refrigerator (Godrej), spectrophotometer (Shimadzu Corporation), Weighing balance, SDS-PAGE assembly (Genei, Banglore) and micropipette (Genei, Banglore) were used to perform necessary experiments.
6
Morphological Analysis of Yunnan Specimens
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All specimens were collected from Yunnan province in China, and mounted according to the methods described by Hodgson and Henderson (2000) . The morphological terminology describing the mounted specimens primarily follows the nomenclature developed by Hodgson (1994) . A Nikon compound microscope was used to examine specimens and an Olympus BH–2 stereoscopic microscope was used to draw illustrations from mounted adult female specimens. Measurements of all characters were recorded in micrometers (μm) or millimeters (mm).
All specimens were deposited in the Northwest A&F University, Yangling, Shaanxi, China (NWAFU
All specimens were deposited in the Northwest A&F University, Yangling, Shaanxi, China (
7
Isolation and Purification of Fungal Cultures
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The fungus from the infected samples was isolated and purified using monoconidial method by streaking out spores on plates containing 2% water agar, pure fungal cultures were obtained by transferring single germinated conidium on potato dextrose agarcontaining antibacterial chloromphenicol (50 μg/ ml) to avoid bacterial contaminations[21 ]. Total 30 cultures were identified by comparing with available literature[22 ] and maintained for further studies. The spores were also verified using compound microscope (Olympus) at different resolutions 4x to 40x.
8
Morphological Identification of BS Causal Agents
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For the morphological identification of BS causal agents, the mycelia, conidia and conidiophores of each isolate (10-day-old) were collected and observed under a compound microscope (Olympus, USA) according to Sivanesan et al.55 and Cardona and Gonzalez56 . Each isolate was screened visually for colour, shape and size.
9
Histological Assessment of EAE Model
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EAE animals and control animals were anesthetized and they were perfused first with saline and then 4% paraformaldehyde and spinal cords were fixed in 10% formalin. To assess infiltration of T cells, H&E stain was performed in paraffin-embedded, 4-μm-thick transverse sections of spinal cord. For assessing demyelination, Luxol Fast Blue was done. Pictures were taken using an Olympus Compound Microscope (Olympus BX-60) with digital camera attached.
10
Whole-mount In Situ Hybridization of c-myb and runx1
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