Immunohistochemical analysis was performed to investigate the expression of HMGA2, Snail, E-cadherin and Vimentin in different grades of human tongue cancer. Briefly, immunohistochemistry (IHC) was performed on the paraffin-embedded human tongue cancer tissue sections. Antigen retrieval was performed in a pressure cooker in citrate solution, pH 6.0, for 15 min, followed by treatment with 3 % hydrogen peroxide for 5 min. Specimens were incubated with antibodies as followed: goat monoclonal antibodies against HMGA2 (1:100, CST), E-cadherin, vimentin, snail (1:100, Santa Cruz, Santa Cruz, CA, USA). For the negative controls, isotype-matched antibodies were applied. The tissue sections were observed under a Zeiss AX10-Imager A1 microscope (Carl Zeiss, Thornwood, NY) and all images were captured using AxioVision 4.7 microscopy software (Carl Zeiss, Thornwood, NY).
Ax10 imager a1 microscope
Manufactured by Zeiss
Sourced in Germany
The AX10-Imager A1 microscope is a high-performance optical microscope designed for advanced imaging and analysis applications. It features a sturdy and reliable construction, with a seamless integration of precision optics and cutting-edge digital imaging technology. The AX10-Imager A1 is capable of delivering clear, high-resolution images, making it a versatile tool for a wide range of scientific and industrial research endeavors.
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Lab products found in correlation
Axiovision 4.7 microscopy software, by Zeiss (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
In situ cell death detection kit, by Roche (1 mentions)
Sp8 scanning confocal microscope, by Leica (1 mentions)
Alexa fluor 488 donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Anti lamp1, by Abcam (1 mentions)
4 6 diamidine 2 phenylindole dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Anti iba1, by Fujifilm (1 mentions)
Anti aβ 6e10, by BioLegend (1 mentions)
Prolong gold, by Thermo Fisher Scientific (1 mentions)
Prolong gold with dapi, by Thermo Fisher Scientific (1 mentions)
L4655, by Merck Group (1 mentions)
Alexa fluor 546 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Filter set 38 he, by Zeiss (1 mentions)
5 protocols using ax10 imager a1 microscope
1
Immunohistochemical Analysis of Tongue Cancer
2
TUNEL Assay for Apoptosis Detection
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This assay was performed using the In Situ Cell Death Detection kit (Roche). Apoptotic cells were also visualized by the terminal deoxytransferase-mediated dUTP-biotin nick end labeling (TUNEL) method. Cells were fixed with 4% paraformaldehyde for 15 min at room temperature, and made permeable with 25 µg/mL of proteinase K for 30 min at 37°C before being incubated with a 1:9 mixture of enzyme solution and labeling solution at 37°C for 2 h. They were washed three times with PBST for 5 min and incubated with peroxidase at 37°C for 30 min. Subsequently, the slides were incubated with 50 µL DAB substrate at room temperature for 10 min. After being washed three times with 1 × TBS, the cells were stained with hematoxylin. The TUNEL-positive cells showed orange-brown nuclei under light ZEISS AX10-Imager A1 microscope.
3
Microglial Immunofluorescence Staining and Imaging
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Microglial cells, plated onto poly-D-lysine-coated cover slips, were fixed with paraformaldehyde (PFA) (4%; 30 min), washed and blocked by incubation with 10% NHS and 0.1% Triton X-100 for 1 h and incubated at 4 °C with the primary antibodies anti-Iba1 (Wako, Japan 1:1000), anti-Aβ 6E10 (Biolegend, USA, 1:500), anti-LAMP1 (Abcam, UK, 1:250) and anti-Arginase (Arg)1 (Santa Cruz, USA, 1:500) followed by the secondary antibodies Alexa Fluor® 594 donkey anti-rabbit IgG (1:1000), Alexa Fluor® 488 donkey anti-mouse IgG (1:1000) and Alexa Fluor® 633 donkey anti-goat IgG (1:1000) and mounted in ProLong®Gold with the nuclear marker 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI; Thermo Scientific, USA). Images (10 fields per experiment, triplicate analysis; × 40 magnification) were acquired with a Zeiss AX10 Imager A1 microscope. Analysis of images was undertaken with ImageJ software; the number of Aβ+ microglia was assessed. Control experiments with each secondary antibody, alone or in combination, were included and images were acquired sequentially on a Leica SP8 scanning confocal microscope for each fluorophore to eliminate cross-signal contamination. Analysis revealed that anti-TLR2 antibody staining of Iba1+ cells was confined to the membrane (see Additional file 1 : Figure S2).
4
Phagocytic Capacity of Microglia
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Uptake of latex beads (1.0 μm mean particle size; L4655, Sigma-Aldrich, Ireland) in a fluorescent yellow-green aqueous suspension was used to analyse phagocytic capacity in microglia isolated from young and aged mice. Cells, prepared as described above, were incubated (4 h) with 0.025% latex beads in fresh cDMEM media. Cells were washed, fixed in 4% PFA, and washed in PBT (PBS + 1% Triton X-100) prior to staining. Cells were blocked (1 h; PBT + 3% BSA), incubated with rabbit anti-Iba1 (19–7141, Wako, Japan 1:1000; overnight; 4 °C), washed, incubated with Alexa Fluor® 546 donkey anti-rabbit IgG (1:1000; 2 h; room temperature, A10040, Invitrogen, USA), and mounted in ProLong Gold with DAPI (P36941, Thermo Scientific, USA). Images (8 fields; 40 × magnification) were acquired with a Zeiss AX10 Imager A1 microscope. Analysis of images was undertaken with the ImageJ software, and the number of latex beads+ microglia was assessed.
5
Visualizing Yeast Membrane Proteins
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Single colonies of UCD932 [gar–] and [GAR+], UCD932 Hxt3-GFP [gar–] and [GAR+], and UCCT 1405 Pma1:m:Neon:KanMX [gar–] and [GAR+] were inoculated into 10 mL of YPD liquid and grown overnight on a rotary drum at 25 °C. Cultures were reinoculated and grown until mid-log phase/exponential phase (∼0.4A600 nm). Stationary cells were taken from the original overnight cultures (∼1A600 nm). Wet mounts were prepared and photographed with the Carl Zeiss AX10 Imager. A1 Microscope (Carl Zeiss, Oberkochen, Germany), Zeiss EX Plan-NEOFLUAR 100X oil immersion lens, with the Zeiss 38 HE Filterset (Ex:440/Em:550). Images were exported in the.tiff format using the Zeiss AxioVision acquisition software version 4.8 from Carl Zeiss. Similar exposure times were used for all images being compared.
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