Macrocolonies were visualized at 10-fold magnification with a Stemi 2000-C stereomicroscope (Zeiss; Oberkochen, Germany). Digital photographs were taken with an AxioCam ICC3 digital camera coupled to the stereomicroscope, which was operated using AxioVision 4.8 software (Zeiss).
Axiocam icc3
Manufactured by Zeiss
Sourced in Germany, United States
The AxioCam ICc3 is a digital camera developed by Zeiss for microscopy applications. It features a high-resolution CMOS sensor and is designed for capturing images of microscopic samples with precision and clarity.
Automatically generated - may contain errors
Lab products found in correlation
Axiovision 4, by Zeiss (9 mentions)
Stemi 2000 c, by Zeiss (4 mentions)
Axioskop 40, by Zeiss (4 mentions)
Axio imager a1, by Zeiss (4 mentions)
Axioplan microscope, by Zeiss (3 mentions)
Axio observer a1, by Zeiss (3 mentions)
Axioscope, by Zeiss (2 mentions)
Axio imager m1, by Zeiss (2 mentions)
Efficient navigation zen , by Zeiss (2 mentions)
Hbo 100, by Zeiss (2 mentions)
Axiovision software, by Zeiss (2 mentions)
Dig rna labeling kit, by Roche (2 mentions)
Imager z2, by Zeiss (2 mentions)
Axio imager m1 microscope, by Zeiss (2 mentions)
Stemi sv11 microscope, by Zeiss (2 mentions)
Plan apochromat 100 1.4 oil ph3 objective, by Zeiss (2 mentions)
Hxp 120 c lightning unit, by Zeiss (2 mentions)
Sytox orange, by Thermo Fisher Scientific (2 mentions)
Axio imager m2 microscope, by Zeiss (2 mentions)
Axio scope a1 optical light microscope, by Zeiss (2 mentions)
72 protocols using axiocam icc3
1
Macrocolony Visualization via Stereomicroscope
2
Characterization of Basidiobolus Strains
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A total of 12 Basidiobolus strains was used for morphological, physiological, and sequence-based characterization (Table 1 ). Nine Basidiobolus spp. isolates were obtained in a previous study [12 (link)] analyzing reptile feces from a suburban area in Pietermaritzburg (KwaZulu Natal, South Africa). Additionally, two isolates (Cla6, C3-1) were obtained from gecko feces collected in Clarens (Free State, South Africa) and identified as described previously [12 (link)]. The type strain Basidiobolus microsporus DSM 3120 was used for comparison. All isolates were kept as living cultures on non-selective media such as Sabouraud Dextrose, Nutrient, and Wort Agar at ambient temperature with regular sub-culturing for maintenance. All microscopic analyses were done using a Zeiss Axio Scope with an Axiocam ICc3. Images were taken using Zen 2.3 blue edition and assembled with Affinity Photo (version 1.8.5.703).
3
Histological Tissue Analysis of Liver
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Small pieces taken from the same liver section were immersed in 10% buffered formalin. Formalin-fixed livers were paraffin embedded and sections measuring 3–5 μm were stained with hematoxylin. The stained tissue sections were viewed under bright field microscopy at 200× magnification. An AxioCam ICc3 camera (Zeiss, Thornwood, NY, USA) was used to take the pictures.
4
Histological Analysis of Calvaria Samples
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After µCT analysis, the calvarias were harvested and fixed in 10% buffered formalin, decalcified in 4% ethylenediaminetetraacetic acid (Merck Millipore, Darmstadt, Hesse, Germany), washed and dehydrated in alcohol. The samples were embedded in paraffin (Sigma-Aldrich), cut with 5 μm thickness and stained with hematoxylin and eosin (Neon, Suzano, SP, Brazil). The images were analyzed and acquired using a light microscope Axioskop 40 (Carl Zeiss, Oberkochen, BW, Germany) coupled to a digital camera Axiocam Icc3 (Carl Zeiss).
5
Visualization of Nanoparticle Distribution in Intestinal Mucosa
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The tissue distribution of nanoparticles in the gastrointestinal mucosa was visualized by fluorescence microscopy. For that purpose, 25 mg of Lumogen® F Red-labeled nanoparticles were orally administered to rats as described above. Two hours later, animals were sacrificed by cervical dislocation and the guts were removed. Ileum portions of 1 cm were collected, cleaned with PBS, stored in the tissue proceeding medium OCT™ and froZEN at −80 °C. Each portion was then cut into 5-µm sections on a cryostat and attached to glass slides. Finally, these samples were fixed with formaldehyde and incubated with DAPI (4′,6-diamidino-2-phenylindole) for 15 min before the cover assembly. The presence of both fluorescently loaded poly(anhydride) nanoparticles in the intestinal mucosa and the cell nuclei dyed with DAPI were visualized in a fluorescence microscope (Axioimager M1, Zeiss, Oberkochen, Germany) with a coupled camera (Axiocam ICc3, Zeiss, Oberkochen, Germany) and fluorescent source (HBO 100, Zeiss, Oberkochen, Germany). The images were captured with the software ZEN (Zeiss, Oberkochen, Germany). As control, a suspension of Lumogen® F Red 305 was administered.
6
Embryo Whole Mount Imaging
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Embryos were whole mounted in slide chambers with 0.6% low-melting poing agarose. To image stained embryos after ISH, images were captured using a 10× objective on an AxioImager Z1 (Zeiss) compound microscope with an Axiocam ICC3 color camera (Zeiss). Images in multiple focal plans were captured individually and combined using the Extended Focus module within Axiovision software (Zeiss). For confocal imaging, embryos were mounted in 0.6% low-melting point agarose and imaged using a 10×, 20×, or 40× objective on a Nikon A1R confocal microscope. Denoising (Nikon Elements software) was performed for some of the images with weak signal to reduce noise. Images were assembled in Adobe Photoshop CS6 software package. Non-linear level adjustment was used to increase contrast and reduce background. In all cases, images of control and experimental embryos were adjusted similarly.
7
Adipocyte Quantification and UCP1 Immunohistochemistry
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The number of adipocytes was counted in hematoxylin-eosin-stained tissue sections of rWAT. Results were expressed as the number of adipocytes per area (mm2). For immunohistochemistry analysis, serial sections of rWAT fixed samples were incubated with normal goat serum 2% in PBS pH 7.3 to block unspecific sites, and then with primary rabbit polyclonal UCP1 antibody (GeneTex International Corporation; Irvine, CA, USA) diluted 1:200 in PBS overnight at 4 °C. Sections were then incubated with biotinylated anti-rabbit IgG secondary antibody (Vector Laboratories; Burlingame, CA, USA) diluted at 1:200, and finally with ABC complex (Vectastain ABC kit, Vector; Burlingame, CA, USA). Peroxidase activity was revealed with Sigma Fast 3,3TM-diaminobenzidine (Sigma-Aldrich; Madrid, Spain) as substrate. Sections were counterstained with hematoxylin and mounted in Eukitt (O. Kindler; Freiburg, Germany). Images were acquired with a Zeiss Axioskop 2 microscope equipped with an AxioCam ICc3 digital camera and AxioVision 40V 4.6.3.0 software (Carl Zeiss).
8
Whole-Mount In Situ Hybridization in Transparent Zebrafish Embryos
9
Histological Analysis of Tissue Constructs
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Representative samples from constructs and brain, lung, heart, liver, kidney, gut, spleen, lymph node, bone marrow were fixed in 4% buffered formalin (Panreac Quimica, Barcelona, Spain) for 48 h. Constructs and bone marrow were additionally decalcified in a formic-acid-based commercial solution (TBD-2, Thermo, Madrid, Spain) for 12–16 h. Samples were then washed, processed and paraffin-embedded. Sections were obtained and stained with hematoxylin and eosin (H&E) for standard histological analyses. To study the presence of human osteocalcin producer cells, a standard indirect ABC immunohistochemical staining was performed, using a specific polyclonal rabbit human anti-osteocalcin antibody (LsBio, Seattle, WA, USA) with a commercial kit EnVision FlexTM, (Dako, Carpinteria, CA, USA)). All samples were evaluated with a conventional light microscope (Axio Scope AX10, Zeiss, Oberkochen, Germany), with attached digital camera (Axio Cam Icc3, Carl Zeiss, Jenna, Germany).
10
Sporangia and Gametangia Formation Assay
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Formation of sporangia was induced by submersing two 12–15 mm square discs cut from the growing edge of a 2–4-d-old V8A colony in a 90-mm-diameter Petri dish in non–sterile soil extract (50 g of oak forest soil in 1 000 mL of distilled water, filtered after 24 h) [39 (link)]. The Petri dishes were incubated at 20 ºC, and natural daylight and the soil extract changed after ca 6 h [74 (link)]. Shape, type of apex, caducity, pedicels and special features of sporangia were recorded after 24–48 h. For each isolate, 50 sporangia were measured at ×400 using a compound microscope (Zeiss Imager.Z2), a digital camera (Zeiss Axiocam ICc3) and biometric software (Zeiss ZEN).
The formation of gametangia (oogonia and antheridia) and their characteristic features were examined after 21–30 d growth at 20 °C in the dark on V8A. For each isolate, 50 oogonia, oospores and antheridia chosen at random were measured under a compound microscope at ×400. The oospore wall index was calculated according to [117 ].
The formation of gametangia (oogonia and antheridia) and their characteristic features were examined after 21–30 d growth at 20 °C in the dark on V8A. For each isolate, 50 oogonia, oospores and antheridia chosen at random were measured under a compound microscope at ×400. The oospore wall index was calculated according to [117 ].
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