Teneral (newly eclosed and unfed adults) G. m. morsitans males were provided an infectious blood meal containing 8x106 BSF T. congolense (VAT TC13) per ml of blood in their first blood meal. After the first infectious blood meal, the flies were maintained on normal blood for the duration of the study. Uninfected control flies were maintained on normal blood only. Twenty-eight days post-challenge (dpc), all flies were dissected 72 h after their last blood meal. Infection status of the PB (defined here as labrum, hypopharynx and thecal bulb) was microscopically determined on Zeiss Axiostar Plus Light microscope at 400x. To dissect the PB, mouth parts were detached from the head and two needles (one in each hand) were used to tease apart the labrum and hypopharynx from the labium. The labium was then detached from the labrum and hypopharynx at the junction of the thecal bulb. This left the labrum, hypopharynx and thecal bulb attached together. Infected labrum and hypopharynx were snap frozen in liquid nitrogen and stored at -80°C until use. In the current study, a total of 7.8% (284/3655) of parasite challenged tsetse had T. congolense infections in the PB. All infected PBs, as well as an equal number of PBs dissected from age-matched uninfected control flies, were divided into two independent biological replicates, each of which contained 130 probosces for subsequent analysis.
Axiostar plus light microscope
Manufactured by Zeiss
Sourced in Germany
The Axiostar Plus is a light microscope designed for routine microscopy applications. It features bright-field illumination and offers a range of magnification options. The Axiostar Plus is suitable for various educational and entry-level laboratory settings.
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Lab products found in correlation
Coverslip, by Thermo Fisher Scientific (2 mentions)
Pc 1049 powershotg5, by Canon (2 mentions)
Nbt bcip, by Roche (1 mentions)
Smz800, by Nikon (1 mentions)
Moticam 5 mp, by Motic (1 mentions)
Anti mouse cd68, by Bio-Rad (1 mentions)
Neubauer chamber, by Zeiss (1 mentions)
Axio scope a1, by Zeiss (1 mentions)
3 3 diaminobenzidine dab substrate, by Merck Group (1 mentions)
Mouse anti human cx43 primary antibodies, by Merck Group (1 mentions)
Goat anti mouse horseradish peroxidase secondary antibody, by Immunotools (1 mentions)
3 3 diaminobenzidine dab liquid substrate system, by Merck Group (1 mentions)
Axio vision le rel 4, by Zeiss (1 mentions)
Axiocam digital camera, by Zeiss (1 mentions)
Paraplast, by Merck Group (1 mentions)
18 protocols using axiostar plus light microscope
1
Tsetse Fly Infection by Trypanosoma congolense
2
Immunohistochemical Analysis of RUNX2 and ADAM17 in MC3T3-E1 Cells
3
Alkaline Phosphatase Staining of MC3T3-E1 Cells
4
Morphological and Anatomical Analysis of Cactus Areoles
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We used young cladodes (smaller than 12 cm long) of parental plants. We removed three areoles from one young cladode of each of three plants (n = 9) using a razor blade and fixed them with FAA solution (3.7% formaldehyde, 48% ethanol, 5% acetic acid, in deionized water) for 72 h according to Ruzin [88 ]. We washed the tissues three times with water and preserved them in GAA solution (25% v/v glycerol, 50% v/v ethanol, 25% v/v water). For morphological description, we observed and characterized areoles using a Nikon SMZ800 stereomicroscope (Nikon Instruments Inc., Japan) and documented images with a Moticam 5MP (Motic Asia, Hong Kong). For anatomy, we sectioned the areoles, prepared them for transversal sections and processed them for standard paraffin embedding protocol using Paraplast Plus (McCormick Scientific, St. Louis, MO, USA) following Ruzin [88 ]. We obtained transversal (8 μm) and serial sections with a rotary microtome (American Optical Company, USA) and stained them with safranin O solution (0.05% safranin O C.I. 50240 in 3% NaCl in water) and fast green FCF solution (0.12% fast green FCF C.I. 42053 in 95% ethanol) following Zavaleta-Mancera et al. [89 ]. We observed the sections with a Axiostar Plus light microscope (Carl Zeiss, Germany) and captured images with a Moticam 5MP camera (Motic Asia, Hong Kong). We documented the procedure digitally [90 ].
5
Immunohistological Analysis of Frontal Cortex
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After pain test on day 4, mice were anaesthetised, perfused successively with cold PBS and 4% PBS-paraformaldehyde via cardiac puncture. Brains were then removed, immersed in 4% PBS-paraformaldehyde and kept for 24 h at 4 °C. Thereafter, tissues were processed using ascending grades of ethanol, xylene (for clearing) and paraffin wax (for embedding). Frontal cortex was sectioned (5µm; MK 1110 rotary microtome) and stained with Cresyl fast violet (CFV) and haematoxylin and eosin (H&E). Also, immunohistochemistry was carried out on the sections of frontal cortex using anti-mouse-CD68 (1:100; BIO-RAD, United Kingdom). Basically, sectioned tissues were deparaffinised, underwent antigen retrieval and subjected to CD68 antibody interaction. Photomicrographs of varied stained tissues were captured using the Zeiss Axiostar plus light microscope.
6
Histological Analysis of Liver Tissue
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Liver tissues were fixed overnight at room temperature in 4% formaldehyde and embedded in paraffin. Sections (8 μm thick) were stained with hematoxylin-eosin and were mounted on glass slides. The stained sections were viewed with an Axio-Star Plus light microscope (Carl Zeiss, Gottingen, Germany). Liver sections were stained with CD68 and XBP-1 antibodies (Santa Cruz).
7
Intestinal Morphology Assessment in Broiler Chickens
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Intestinal morphology was assessed by histology at 21 and 34 d. On 21 d, 2-cm segments of duodenum, jejunum and ileum were collected from 6 birds randomly selected from both the control group and from the group fed the DON diet from 1 to 21 d. On 34 d, 6 birds from each of the 5 treatments were randomly selected and intestinal segments collected. Tissues were fixed in 10% neutral buffered formalin and then transferred to ethanol until processing. Tissues were paraffin embedded, sectioned at 5-μm thickness and stained with hematoxylin and eosin by Prairie Diagnostic Services Inc. (Saskatoon, SK, Canada). Two cross sections of each tissue were prepared for evaluation. Slides were examined under an Axiostar plus light microscope (Carl Zeiss Microscopy, LLC, NY, USA) and pictures of intestinal epithelial structures were acquired with × 5 objective. Measurements were done on 10 to 16 individual villi and associated crypt per bird. All 3 intestinal regions were evaluated for changes in villus height, crypt depth, villus width, muscularis thickness and villi:crypt ratio.
8
Quantifying Drosophila Larval Hemocytes
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Late wandering third-instar larvae were picked from the walls of the food vial with a pair of forceps and cleaned from food and debris by placing them in a 1X PBS containing petri plate before being transferred to a tissue paper to dry them. The larvae were placed in 10 μl of 1X-PBS that was placed on parafilm strip under a light microscope. Using two pairs of forceps the larval cuticle was pierced and hemolymph was released. The larva was left to bleed for 20 secs. The hemolymph bleed was pipetted and mixed with an equal volume of trypan blue. A total of 10 μl of the mix was placed in a Neubauer chamber (Buerker-Turk, Marienfeld, Germany) with a coverslip attached. The Neubauer chamber was placed under an Axiostar plus light microscope (Zeiss, Oberkochen, Germany) and number of cells in each of the four quadrants was noted down. Hemocyte number was then reported as hemocytes per milliliter of bleed. The average number of hemocytes was obtained from three different biological replicates (N=3) with ten third-instar larvae for each replicate. The total number of larvae used in each condition is (n=30).
9
Intestinal Morphology and Goblet Cell Analysis
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Ileal tissue samples were obtained 15 cm from the ileocecal junction and immediately fixed in 10% buffered formalin. The fixed intestinal segments were embedded in paraffin and sectioned for intestinal morphology (Prairie Diagnostic Services, Saskatoon, SK). Briefly, sections of the tissue were deparaffinized in xylene, rehydrated and stained with haematoxylin and eosin. Slide images were measured at 10 × magnification using an Axio Star Plus light microscope (Axio Scope A1; Carl Zeiss Gottingen, Germany). The villus height and crypt depth of each tissue were measured using the AxioVision Rel 4.8 software (Carl Zeiss Canada Ltd., Toronto, ON) on a minimum of 10 well-oriented villi and their corresponding crypts per sample. Goblet cell counts were determined by preparing tissue samples as indicated above and staining with Alcian Blue and Periodic Acid Schiff as previously described [27 (link)]. The slides were then viewed under a light microscope at 10 × magnification (Axio Scope A1; Carl Zeiss Gottingen, Germany). For each tissue sample, 10 well-oriented villi were selected, and goblet cells were counted in a region within 100 μm length of the villi.
10
Histological Analysis of Liver Tissue
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Liver tissues were fixed overnight at room temperature in 4% formaldehyde and embedded in paraffin. Thereafter, the tissue sections (8 μm thick) were stained with hematoxylin and eosin (H&E) and mounted on glass slides. The stained sections were viewed under an Axiostar Plus light microscope (Carl Zeiss, Gottingen, Germany).
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