For observation of the penetration and invasive growth in rice cells, conidial suspensions (1 × 105 spores ml−1) of Guy11, mutants and complement strains were injected into rice leaf sheath. The inner epidermises of infected sheaths were harvested at different hours postinoculated by wild‐type and mutant strains and observed under a microscope. The complement strains with GFP fluorescent protein in the infected rice cells were observed using fluorescence microscopy (Zeiss Axio Observer A1 inverted microscope, ×63 magnification oil objective). Observation of effector secretion was performed as in our previous work (Qi et al., 2016 ). Briefly, for construction of Avr‐Pia:GFP, Avr‐Piz‐t:GFP and Bas4:GFP were transformed into Guy11, the ∆Moswa2 mutant and the ∆Mosec24‐2 mutant. The conidial suspensions were collected and followed the above infection assay. GFP fluorescence was observed using a fluorescence microscope (Zeiss Axio Observer A1 inverted microscope, ×63 magnification oil objective).
Axio observer a1 inverted microscope
Manufactured by Zeiss
Sourced in Germany
The Axio Observer A1 is an inverted microscope designed for a variety of applications. It features a stable and compact design, supporting a range of contrast techniques and objectives. The Axio Observer A1 provides a reliable platform for various imaging and analysis tasks.
Automatically generated - may contain errors
Lab products found in correlation
Mrc5 camera, by Zeiss (4 mentions)
Pbs buffer, by Thermo Fisher Scientific (4 mentions)
Dmem high glucose media, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (4 mentions)
Pen strep, by Thermo Fisher Scientific (4 mentions)
Ultra low attachment 96 well plate, by Corning (3 mentions)
Nile red oxazone, by Merck Group (2 mentions)
Agilent 6230 lc tof mass spectrometer, by Agilent Technologies (2 mentions)
Tribute peptide synthesizer, by Protein Technologies (2 mentions)
Sp5 confocal fluorescence microscope, by Leica (2 mentions)
Facsaria cell sorter, by BD (2 mentions)
Rat tail collagen type 1, by BD (2 mentions)
Transwell insert, by Merck Group (2 mentions)
Hiscript 2 reverse transcriptase, by Vazyme (2 mentions)
Methylcellulose, by Merck Group (2 mentions)
Prism 6, by GraphPad (2 mentions)
Duolink in situ red starter kit mouse rabbit, by Merck Group (2 mentions)
Src 3, by Merck Group (2 mentions)
Pfkfb4, by Merck Group (2 mentions)
Pfkfb4, by OriGene (2 mentions)
52 protocols using axio observer a1 inverted microscope
1
Rice Cell Penetration and Invasion
2
Visualizing Glycogen and Lipids in Fungal Structures
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The glycogen metabolism in the germinating conidia and appressoria of strains were visualized by staining these tissues with glycogen staining solution containing 60 mg/ml KI and 10 mg/ml I2 [77 ]. Once the samples become yellowish-brown, the glycogen deposits can be visualized in bright field optics with Zeiss Axio Observer A1 inverted microscope.
The lipid droplets in the germinating conidia and appressoria of strains were visualized by staining these tissues with a Nile red solution consisting of 50 mM Tris/maleate buffer (pH 7.5) and 2.5 mg/ml Nile red Oxazone (9-diethylamino-5H-benzo-a-phenoxazine-5-one, Sigma) [10 (link), 55 (link), 78 (link)]. After 3 min incubation, the lipid droplets in the conidia and appressoria began to fluoresce and were observed under Zeiss Axio Observer A1 inverted microscope
The lipid droplets in the germinating conidia and appressoria of strains were visualized by staining these tissues with a Nile red solution consisting of 50 mM Tris/maleate buffer (pH 7.5) and 2.5 mg/ml Nile red Oxazone (9-diethylamino-5H-benzo-a-phenoxazine-5-one, Sigma) [10 (link), 55 (link), 78 (link)]. After 3 min incubation, the lipid droplets in the conidia and appressoria began to fluoresce and were observed under Zeiss Axio Observer A1 inverted microscope
3
Histological analysis of spinal motion segments
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15 μm sections from L5‐L6 motion segments were processed using H&E according to standard protocols. In short, sections were post fixed for 15 min with 4% PFA, washed with 1X PBS, and stained with hematoxylin. Following hematoxylin staining, sections were washed with 1X PBS, differentiated using acid alcohol, blued with a sodium acetate solution, dehydrated using an alcohol gradient, counterstained with eosin, dehydrated with xylenes, and mounted using Permount (Fisher Scientific SP15‐100). Three motion segment sections from each animal were stained using the aforementioned process and each motion segment was imaged by collecting and stitching two images taken with a 10X objective on a Zeiss Axio Observer A1 Inverted Microscope (Carl Zeiss Microscopy, Inc.).
4
Tube Formation Assay for Neonatal LECs
5
Meridianin C-Induced Macropinocytosis Monitoring
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To monitor the functionality of meridianin C‐induced macropinocytosis (macropinosome formation/internalization), 0.25 × 105 YD‐10B cells/mL were seeded on coverslips and treated with meridianin C (1 μM) and/or FITC‐dextran (0.5 mg/mL) in the presence or absence of amiloride (4 mM) for 4 h. The cells were washed twice with PBS and mounted onto microscopic glass slides using Permafluor aqueous mounting media (Thermo Scientific, Waltham, MA, USA) media. Bright field and fluorescence were observed using a Zeiss AxioObserver.A1 inverted microscope (Carl Zeiss, Germany) and images acquired using Zen 2 software (Carl Zeiss). Fluorescent intensity was quantified using Image‐J software.
6
Small Molecule and Peptide Synthesis Protocol
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Chemical reagents for small molecule and peptide synthesis were purchased from various vendors and used as received. The phospholipids were purchased from Avanti Polar Lipids (Alabaster, Al). PBS buffer, DMEM/High glucose media, RPMI 1640 media, and Pen/Strep were purchased from Thermal Scientific (Amarillo, TX). The Gram-positive bacteria (B. subtilis (ATCC 663) and S. aureus (ATCC 6538)) were purchased from Microbiologics (Cloud, MN) as lyophilized cell pellet. E. coli (BL 21) was a gift from the lab of Professor Mary F. Roberts at Boston College. NMR data of the small molecules were collected on a VNMRS 500 MHz NMR spectrometer. MS data were generated by using an Agilent 6230 LC TOF mass spectrometer. Peptide synthesis was carried out on a Tribute peptide synthesizer from Protein Technologies. The fluorescence anisotropy experiments were performed by using a SpectraMax M5 plate reader. Fluorescence images were taken on a Zeiss Axio Observer A1 inverted microscope. Confocal images were taken on the Leica SP5 confocal fluorescence microscope housed in the Biology Department of Boston College. Flow cytometry analyses were carried out on a BD FACSAria cell sorter also housed in the Biology Department of Boston College.
7
Neutralization Assay for CedPV Glycoprotein Antibody
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Murine monoclonal antibody (mAb) m14F3, is a CedPV soluble G specific mAb of IgG2 subclass that was developed using standard hybridoma generation techniques by immunization of BalbC mice with purified, recombinant soluble CedPV G glycoprotein produced from a human 293F stably-expressing cell line. HeLa-USU cells were seeded at a density of 5 × 104 cells/well in 96-well cell culture plates. The next day, the indicated dilutions of mAb m14F3 were incubated with equal volumes of rCedPV-GFP (MOI: 0.1) for 1 h at 37 °C. was removed from the cells and 100 μl of virus or the antibody-virus mixture was added to the wells in triplicate and allowed to incubate for 1 h at 37 °C. After the incubation, the antibody-virus mixture and virus only was removed and all cells were washed once with DMEM-10. Fresh DMEM-10 with the varying concentrations of m14F3 were added to the corresponding wells and incubated for 48 h at 37 °C. GFP foci were counted with a Zeiss Axio Observer A1 inverted microscope using the 5X objective. The percent neutralization was determined based on the presence of GFP foci in replicate wells at each antibody concentration and calculated based on the reduction in the average number of foci per well to the average number of foci observed in the no antibody control, multiplied by 100 [70 (link)].
8
Quantitative Phase Imaging of Cell Lines
9
Histological Evaluation of Decellularized DSMG
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Histological analysis was carried out immediately after decellularization to confirm complete decellularization of the prepared DSMG. This DSMG was then fixed in ice-cold paraformaldehyde and dehydrated in graded ethanol. Next, 5-μm embedded paraffin sections were stained with hematoxylin and eosin (H&E) and evaluated under light microscopy (Zeiss®, Axio Observer A1 inverted microscope).
10
Immunofluorescent Muscle Fiber Typing
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