Cell growth, morphology, and viability were controlled during cell culture with a phase contrast microscope (Leica DM IL LED) with LAS EZ software (Leica DM 750 Microsystems, Switzerland). The cell viability during force experiments was controlled with a digital camera (uEye capture device filter with camera model UI148XLE-C, Obersulm, Germany) connected to the AFM instrument. Typically, cells started gradually to detach and die after 2 hours of experiments, so the measurement time was always kept under 2 h or 1.5 h for HepG2 and WA07 cells, respectively. In addition to the visual observation of the cell morphology commonly used in AFM force spectroscopy studies to monitor cell state, we also checked the cell viability after the force measurements by the Trypan Blue exclusion test with cell fixation for adherent cells, a protocol provided by Perry et al.62 . Briefly, cells were dipped into Trypan Blue Solution (1:5 dilution, Trypan Blue Solution, 0.4%, GibcoTM, 15250-061) and washed with 1 × DPBS solution. After fixation in 4% paraformaldehyde solution for 10 min the cells were washed twice and mounted on the objective glass with ProLong® Gold Antifade reagent (Invitrogen, P36934). The imaging of the fixated cells was performed with a Leica phase contrast microscope (Leica DM750) with LAS EZ software.
Dm750
Manufactured by Leica
Sourced in Germany, Switzerland, China, United States, Japan, Italy
The DM750 is a microscope designed for routine laboratory applications. It features a high-performance optical system and an ergonomic design, providing reliable performance and user comfort.
Automatically generated - may contain errors
Lab products found in correlation
Icc50 hd, by Leica (14 mentions)
Icc50w, by Leica (7 mentions)
Icc50, by Leica (4 mentions)
Icc50w digital camera, by Leica (3 mentions)
Icc50 e, by Leica (3 mentions)
Dab peroxidase substrate kit, by Vector Laboratories (3 mentions)
Gar hrp, by Microtech (3 mentions)
Dmc2900, by Leica (3 mentions)
Dfc295, by Leica (3 mentions)
Dm2500, by Leica (3 mentions)
Tp1020, by Leica (3 mentions)
Application suite, by Leica (2 mentions)
Application suite 4, by Leica (2 mentions)
Ab32420, by Abcam (2 mentions)
Anti rad51c, by Thermo Fisher Scientific (2 mentions)
Image pro plus, by Media Cybernetics (2 mentions)
Mc170 hd, by Leica (2 mentions)
Entellan, by Merck Group (2 mentions)
Fei quanta 200 feg, by Thermo Fisher Scientific (2 mentions)
Las core v4, by Leica (2 mentions)
288 protocols using dm750
1
Cell Viability Monitoring in AFM Experiments
2
Histological Analysis of Murine Cecum
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The cecum tissues of mice were stained with hematoxylin and eosin (H&E) (Servicebio, Wuhan, China) and then observed using a Leica DM750 software (Leica DM750, Leica, Beijing, China) to evaluate the morphological changes in the cecum.
3
Ocular measurements of parasitoid wasps
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All measurements are in millimeters and taken initially from micrometer divisions directly fitted in the eye piece of a Leica S8APO stereo zoom trinocular microscope at 80× for card mounted specimens or at 100× using a Leica DM750 phase contrast microscope for slide mounted parts and finally converted into millimeters. Images of card mounted specimens were captured using a Leica M205C stereo zoom trinocular microscope with a DMC2900 camera, and those of slide mounted parts using a DFC295 camera attached to a Leica DM750 phase contrast microscope.
The following abbreviations are used:
AOL = Minimum distance between a posterior ocellus and the anterior ocellus.
F1, F2, etc. = Funicle segments 1, 2, etc.
OCL = Minimum distance between a posterior ocellus and the occipital margin.
OOL = Minimum distance between a posterior ocellus and the corresponding eye margin.
POL = Minimum distance betweenposterior ocelli.
The following acronyms are used for the depositories:
EDAU = Entomology Department, Annamalai University, Chidambaram, India.
NBAIR = National Bureau of Agricultural Insect Resources (Formerly NBAII), Bangalore, India.
The following abbreviations are used:
AOL = Minimum distance between a posterior ocellus and the anterior ocellus.
F1, F2, etc. = Funicle segments 1, 2, etc.
OCL = Minimum distance between a posterior ocellus and the occipital margin.
OOL = Minimum distance between a posterior ocellus and the corresponding eye margin.
POL = Minimum distance betweenposterior ocelli.
The following acronyms are used for the depositories:
EDAU = Entomology Department, Annamalai University, Chidambaram, India.
NBAIR = National Bureau of Agricultural Insect Resources (Formerly NBAII), Bangalore, India.
4
Histological and Immunohistochemical Analysis of Liver Samples
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Liver samples were collected and fixed in 4% paraformaldehyde overnight, then embedded in paraffin and cut into 5μm sections for hematoxylin and eosin (HE) staining. The histological changes were observed under an optical microscope (DM750, Leica, Shanghai, China), and scored according to Suzuki’s criteria, which are detailed in a previous report [11 (link)]. For immunohistochemistry staining, liver sections were deparaffinized in xylene, hydrated in gradient alcohol, and antigen-repaired in citrate buffer (pH = 6.0). After that, liver sections were blocked with goat serum at 37 °C and then incubated with primary antibodies, including CD68 (Abcam, Shanghai, China) and F4/80 (Abcam, Shanghai, China), overnight at 4 °C. The sections were incubated with HRP-conjugated antibody, followed by staining with 3,3′-diaminobenzidine (DAB), and counterstained with hematoxylin. After viewing and imaging under a brightfield microscope (DM750, Leica, Shanghai, China), the IHC positive staining areas were quantified using ImageJ software (NIH, Bethesda, MD, USA).
5
Histopathological Analysis of Control and Case Samples
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The specimens were received in the pathology department as 2 samples from each patient in separate bottles as sample A-Control and sample B-Case/test. Routine processing for histopathological examination were done, embedded in paraffin wax and 5 micron thick sections were stained with Hematoxylin and Eosin stain. Histomorphological observation was done under Leica-DM 750 image analysis microscope. The following observations were made in both control and case samples.
6
Macro- and Micro-Morphological Characterization of Fungal Strains
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For macro-morphological characterization, strains were cultured on PDA, Malt Extract Agar (DifcoTM, Sparks, MD, USA) supplemented with 10% NaCl (w/v) (MEA 10%), Leibniz Institute DSMZ—German Collection of Microorganisms and Cell Cultures 372-Halobacteria medium amended with 10% NaCl (w/v) (HM 10%), and Dichloran Glycerol Agar (DG 18, Oxoid, Basingstoke, UK) at 25 ± 2 °C for 2 to 6 months [57 (link)]. Morphological traits such as colony diameter, mycelium color, texture, and form, as well as other characteristics, were recorded via direct observation of the cultured media plates. For micro-morphological characterization, PDA (DifcoTM, Sparks, MD, USA) and synthetic low-nutrient agar (SNA), following the recipe by Nirenberg [83 (link)], were used and observed directly with a light microscope (Leica DM750 (Leica, Wetzlar, Germany)), as well as using the slide culture technique, and both were photographed with a Leica ICC50W digital camera (Leica, Wetzlar, Germany). At least 50 measurements per structure were considered. Both strains were deposited and preserved in Micoteca da Universidade do Minho (MUM), Braga, Portugal.
7
Quantifying Interlobular Collagen Fibrosis
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We performed the measurements of the fibrotic septa in the interlobular space on 3 different fields per slide of 3 individual rats per time for each of the treatments. A photomicroscope Leica DM750, camera ICC50W. Heerbrugg, Switzerland and the LAS EZ version 3.4.0 software Leica Microsystems, Switzerland were used to take the pictures and the Axio Vision Rel. 4.6 software. Oberkochen, Germany to make the measurements. We determined the areas of collagen in the interlobular zone as pixels at a total magnification of 100×. Parts that did not show the blue staining pattern characteristic of collagen of Masson’s trichrome staining were excluded. The results were expressed as the total number of positive pixels for collagen.
8
Drosophila Wing Imaging Protocol
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Drosophila wings were mounted using Gary's Magic Mountant, a mixture of Canada balsam and methyl salicylate (4:1 v/v). Images of adult Drosophila wings were taken by Leica DM750 equipped with ICC50E (Leica) using 5× (numerical aperture: 0.15) chroma objectives. Data acquisition was performed using Leica Application Suite, version 4.10.0 (Leica).
9
Reactive Oxygen Species Detection in Plants
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Reactive oxygen species detection was carried out using histochemical staining methods. 3′, 3′-Diaminobenzidine (DAB) and Nitroblue tetrazolium (NBT) were used for detecting hydrogen peroxide and superoxide, respectively. For NBT staining, the seedlings were incubated in a solution of 1/10 MS medium for 10 min and then transferred to a 50 μM NBT solution dissolved in 1/10 MS medium for another 5 min at room temperature. Afterwards the samples were rinsed several times and the roots were irradiated with UV-B for 20 min. Likewise, DAB staining was carried out using the following procedure: the seedlings were incubated in a Tris buffer (pH 5.0) for 10 min and then infiltrated with the DAB solution (final concentration 0.7 mg/ml), for 5 min in a vacuum chamber at room temperature. The samples were washed twice with the Tris buffer. The roots were then treated with UV-B radiation for 20 min. The images of roots were captured with ×10 objective lens of a light microscope Leica DM750 (Solms, Germany). The staining intensity in root apex region was digitized and compared using densitometric method by ImageJ software (ver. 1.43u for Macintosh OSX).
10
Stomatal Aperture Quantification Protocol
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Detached rosette leaves from 4-week old plants were floated in opening buffer (5 mM KCl, 10 mM MES, pH 5.6) for 2 h in the light. Hundred microliters of 50 μM ABA in 2% (v/v) ethanol or 2% (v/v) ethanol (control) was added and the leaves were incubated for 2 h. To measure the response to drought, 4-week-old plants were exposed to 100% humidity for 12 h. Rosette leaves were detached from the plant and incubated on the lab bench for 1 h. Following treatment, the leaves were grinded in opening buffer with a polytron and the homogenate was filtered through nylon cloth (30 μm mesh size). The isolated epidermal fragments were transferred to microscope slides and viewed under the microscope (Leica DM750, Leica, Denmark), 400x magnification.
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