For the microscopy evaluation, bacteria were grown on brain–heart infusion (BHI) agar for 3–5 days at 18°C. The cell morphology of green snow fast‐growing bacteria was studied by examining Gram‐stained cells using widefield microscope Leica DM4 B (Leica Microsystems, Germany) with an objective magnification of ×100 (Harisha, 2007 ). For identifying the optimal growth temperature, bacteria were cultivated on MPA at temperatures 4°C, 10°C, 18°C, 25°C, and 37°C for 10 days with the daily evaluations of the growth by microscopy with widefield microscope Leica DM4 B (Leica Microsystems, Germany) and objective magnification of ×100 (Harisha, 2007 ), the colonies larger than 1.0 mm were visually evaluated. The evaluation of growth was performed according to the score‐based scale where score “0” indicates the absence of growth, score “1–2” indicates a little growth, score “3–4” indicates relatively good growth without single colonies, score “5–6” indicates the growth with the single colonies 0.5–1.0 mm in diameter, score “7–8” indicates the growth when the diameter of colonies was 1.5–2.0 mm, and score “9–10” indicates the growth when the diameter of colonies was higher than 2.5 mm.
Dm4 b
Manufactured by Leica
Sourced in Germany
The Leica DM4 B is a high-performance microscope designed for advanced biological and materials science research. It features a robust and ergonomic design, advanced optics, and a range of customization options to meet the needs of a diverse research community.
Automatically generated - may contain errors
Lab products found in correlation
Dm6 b, by Leica (1 mentions)
Ab39012, by Abcam (1 mentions)
A 21424, by Thermo Fisher Scientific (1 mentions)
Ab15580, by Abcam (1 mentions)
A21429, by Thermo Fisher Scientific (1 mentions)
Ab137321, by Abcam (1 mentions)
Anti fluorescence quenching agent, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Eg1150, by Leica (1 mentions)
Dfc9000, by Leica (1 mentions)
E cadherin 20874 1 ap, by Proteintech (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Dig rna labeling kit, by Roche (1 mentions)
Las af v1, by Leica (1 mentions)
Lsrfortessa, by BD (1 mentions)
Alizarin red s staining solution, by Solarbio (1 mentions)
Spectramax id5, by Molecular Devices (1 mentions)
22 protocols using dm4 b
1
Microscopy Evaluation of Bacteria Growth
2
Cloning and Transient Expression of HvTDF1-GFP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HvTDF1 coding sequence without stop‐codon was amplified and inserted into pCR™8/GW/TOPO™ vector to generate a pENTRY vector (Table S1 ). HvTDF1 coding region with the flanking attL1 and the attL2 region was amplified by Phusion® High‐Fidelity DNA Polymerase (Primers 1857, 1858; Table S1 ) using pCR8GW: HvTDF1‐nonstop ENTRY vector as template. The purified PCR product was used to generate pUbi10pro:HvTDF1‐GFP, the sequence‐confirmed vector was co‐transformed with pBIN‐P19 into Agrobacterium GV3101. A positive Agrobacterium colony was cultured overnight to OD600 0.4–1 with Spectinomycin (100 mg ml−1), Kanamycin (50 mg ml−1) and Rifampicin (50 mg ml−1) in 1 : 1000 dilution, pelleted and resuspended in infiltration buffer and infiltrated into 4‐ to 6‐wk‐old Nicotiana benthamiana Domin, leaves (Cui et al., 2016 (link)). After 48 h, infiltrated leaves were analysed for fluorescence signal expression (Leica DM4B; Leica Microsystems, Milton Keynes, UK); confirmation of protein subcellular localization was conducted by infiltration with 4,6‐diamidino‐2‐phenylindole (DAPI; 10236276001; Sigma‐Aldrich; 10–20 mins; 10 μg ml−1). DAPI and green fluorescent protein (GFP) were observed using a Leica DM4B microscope (DAPI: filter cube LED365; GFP: filter cube LED470).
3
Viability Assay of Pneumococcal Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The changes in viability of penicillin-sensitive S. pneumonia ATCC49619 and penicillin-resistant S. pneumonia 16167 after incubation with PEGylated Nano-BA12K were further assessed using LIVE/DEAD®BacLight™ Bacterial Viability Kit (L7012) after treatment of PEGylated Nano-BA12K, BA, and Penicillin G. Briefly, S. pneumonia cells (~107 CFU/mL) were incubated with tested formulations at 1× MIC for 0.5, 1, 2, 4, 8, and 12 hours. Then the bacterial cells were washed three times with PBS with centrifugation at 3,000 rpm for 10 minutes. Combination of equal volumes of SYTO 9 dye (component A) and propidium iodide (component B) in a microfuge tube was mixed thoroughly. Add 3 µL of the dye mixture for each milliliter of the bacterial suspension, mix thoroughly, and incubate at room temperature in the dark for 15 minutes. After staining, the bacterial cells were rinsed with PBS twice. Subsequently, trap 5 µL of the stained bacterial suspension between a slide and a 20 mm2 coverslip, and the microscope images were captured using an FM (Leica DM4 B; Leica Microsystems, Wetzlar, Germany) at λex (488 nm)/λem (590 nm) for green fluorescence or λex (568 nm)/λem (630 nm) for red fluorescence.
4
Quantifying Podocyte Apoptosis in Kidney Disease
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed, 4 µm thin mice kidney tissue sections were stained using a TUNEL staining kit (#ab66110) according to the manufacturer’s instructions. Further, human podocyte apoptosis was detected in vitro by DAPI staining. Human podocyte cells were grown on the coverslip and treated with or without AGEs, AGEs+DAPT, and AGEs+FPS-ZM1. Next cells were fixed with 4% paraformaldehyde-phosphate buffer saline (PBS) and washed twice with ice-cold PBS. Following these, the cells were permeabilized with 0.1% Triton X-100 in PBS for 10 min at 37°C and stained with 1 mg/mL DAPI dissolved in PBS for 30 min at 37°C. The cells were rinsed twice with ice-cold PBS and fluorescent images were captured from the images acquired using Upright Microscopes Leica DM4 B and DM6 B (Leica Microsystems trinocular). The percentage of condensed nuclei was calculated as the ratio of condensed nuclei to total cells counted, and a minimum of 100 cells/field and at least six fields in each well were counted.
5
Gram Staining and Microscopy Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Gram staining and microscopy evaluation, the isolated bacteria were cultivated on brain heart infusion (BHI)‐agar at 18°C for 1–4 days, depending on the isolate, until the single colony appeared. Gram staining was done following the protocol of the three‐step Gram stain procedure kit (Merck KGaA, Germany). The morphology of Gram‐stained cells was studied by direct examination with the light microscope Leica DM4 B (Leica Microsystems, Germany) under a 100× immersion lens.
6
Histological Analysis of Cartilage Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were fixed in 4% paraformaldehyde/phosphate-buffered saline (PFA/PBS) overnight at 4 °C, embedded in paraffin, and cut into 7 μm-thick sections. The sections were deparaffinized, rehydrated, and then stained with Mayer’s hematoxylin and eosin (H&E). For antigen retrieval, deparaffinized sections were incubated with 5% hyaluronidase in PBS for 30 min at 37 °C for Col2, Col10, and Mmp13 immunostaining. After blocking with 5% bovine serum albumin (BSA), sections were incubated with anti-Col2 (1:1000, #7050; Woodinville Chondrex, WA, USA), anti-Col10 (1:500, LSL, Tokyo, Japan), anti-Mmp13 (1:200, ab39012; Abcam, Cambridge, UK), anti-Pcna (1:1000, #13110; Cell Signaling Technology, Danvers, MA, USA), or anti-Ki67 (1:200, ab15580; Abcam) antibodies at 4 °C for 16 h, washed thrice with PBS, and then incubated with Alexa Fluor 555-conjugated anti-rabbit immunoglobulin G (IgG) (1:500, A21429; Thermo Fisher Scientific) or anti-mouse IgG (1:500, A21424; Thermo Fisher Scientific)28 (link),61 (link). Sample immunoreactivity was analyzed under a fluorescence microscope (Leica DM4 B; Leica Microsystems, Wetzlar, Germany).
7
Immunofluorescence Staining of Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thy-1 + OFs were seeded in 24-well plates on coverslips at approximately 2 × 10 4 per well and cultured overnight at 37 ℃ in an atmosphere of 5% CO 2 . After washing three times with PBS, the cells were fixed with 4% paraformaldehyde for 20 min, permeabilized with PBS containing 0.2% Triton X-100 (PBST) for 30 min and blocked with 3% BSA/PBS for 30 min at room temperature. The cells were then incubated overnight with primary antibodies against fibroblast surface protein (FSP) (SAB4200821, 5 µg/mL, Sigma-Aldrich, Saint Louis, MO, USA) and vimentin (VIM) (ab137321, 1/500 dilution, Abcam) at 4°C. Then the slides were washed with PBS for 5 min again and incubated with Alexa Fluor 488 goat anti-mouse IgG or Alexa Fluor 594 goat anti-Rabbit IgG secondary antibody (Bioss, Beijing, China) at room temperature in the dark for 1 h. All the agents were applied in accordance with the manufacturer's instructions. Nuclei were stained for 10 min with DAPI (Beyotime). After a final wash, an anti-fluorescence quenching agent (Beyotime) was added to mount the coverslips. Images were obtained using Leica DM4B (Leica Microsystems, Brønshøj, Denmark).
8
Scratch Wound Assay for Breast Cancer Cell Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wound healing was used to observe the migration ability of breast cancer cells. A total of 5×104 cells were plated in 6-well plates and cultured until 95% confluency. A plastic 20 µl pipette tip was used to scratch a vertical wound. Detached cells were removed and phase contrast images of the scratched fields were captured at 0 and 24 h. In each group, at least three scratched fields were recorded using an upright light microscope at magnification, ×20 (Leica DM4B; Leica Microsystems, Shanghai, China).
9
Paraffin Embedding and Histological Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All samples were embedded in paraffin using a paraffin-embedding module (Leica EG1150, Leica, Germany) and cut into sections (4 μm thickness) in the longitudinal direction. Microscopic evaluation was performed using a light microscope (Leica DM4 B, Leica, Germany) connected to a digital camera (Leica DFC9000, Leica, Germany). Microscopic evaluation was performed only ex vivo, that is, on resected tissues. Hematoxylin and eosin staining was performed to evaluate the morphology of the tissue samples.
10
Histopathological Analysis of Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histopathological sections were prepared and observed according to our previous study [34 (link)]. A total of six individuals were sampled and fixed in each group, two sections were made for each individual, and three non-contiguous tissue sections were randomly analyzed for each section. Overall, a total of six pathology sections were observed per individual in each group using a light microscope (LEICA DM4 B, Leica, Wetzlar, Germany). The magnification of all sections was approximately 40×.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!