The inner pore size distribution and porosity of microspheres were determined by Mercury Injection Apparatus (AutoPore IV 9500, Micromeritics). For the glass transition temperature of mixture (PLA and PELA) measurement, lyophilized mixture was analyzed by differential scanning calorimetry (DSC Q2000). The process of antigen penetrating into microcapsule inner structures was observed by CLSM TCS SP8 (Leica, Germany).
Tcs sp8 clsm
Manufactured by Leica camera
Sourced in Germany
The TCS SP8 CLSM is a confocal laser scanning microscope manufactured by Leica. It is designed to provide high-resolution imaging of biological samples. The microscope utilizes multiple laser lines to excite fluorophores within the sample and captures the emitted fluorescence using sensitive detectors. The system offers advanced features for three-dimensional imaging, time-lapse experiments, and image analysis.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Autopore 4 9500, by Micromeritics (1 mentions)
D7200, by Nikon (1 mentions)
Su8010, by Hitachi (1 mentions)
Ab32575, by Abcam (1 mentions)
Normal horse serum, by Vector Laboratories (1 mentions)
Alexa fluor 488 fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions)
Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions)
Influx flow cytometer, by BD (1 mentions)
Ivis spectrum in vivo imaging system, by PerkinElmer (1 mentions)
Cfx connect software, by Bio-Rad (1 mentions)
Jem 1200ex tem, by JEOL (1 mentions)
Chemidoc xrs imaging system, by Bio-Rad (1 mentions)
Chirascan circular dichroism spectrometer, by Applied Photophysics (1 mentions)
Uv 3600 spectrophotometer, by Shimadzu (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Las x version, by Leica camera (1 mentions)
Imaris software, by Oxford Instruments (1 mentions)
13 protocols using tcs sp8 clsm
1
Characterization of Microsphere Porosity
2
Sunflower Seed Microstructure Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A suitable quantity of sunflower seeds was neatly arranged, and the appearance was photographed and sampled using a camera (D7200, Nikon, Tokyo, Japan). The microstructures of sunflower seeds were examined using Scanning Electron Microscopy (SEM, Hitachi SU8010, Tokyo, Japan). The shell was carefully removed without causing any damage to the raw materials of sunflower seeds, and the method of Xiang et al. [20 (link)] was slightly modified. Then it was observed under SEM following gold spraying with a magnification of ×500. The microscopic morphology of the semi-thin section cells of the sunflower seeds was observed by Confocal Laser Scanning Microscopy (CLSM TCS-SP8, Leica, Germany) [21 (link)]. The sunflower seeds were cut to an appropriate size, then moistened in the plant fixing solution, and semi-thinly sliced using a microtome. Semi-thin slices of sunflower seeds were stained with 0.2% fluorescein isothiocyanate and 0.1% Nile red, respectively. After a 3 min staining period in the absence of light, the excess staining medium on the surface was rinsed off with ×1 phosphate buffer, and the structure of sunflower seed cells was observed with a magnification of the original field of view ×63.
3
Immunofluorescent Analysis of ACTA2 and TNMD
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples (n = 5–6) were fixed with 10 (v/v)% formalin and permeabilized with 0.1 (v/v)% X-100 Triton (ThermoFisher Scientific, Waltham, MA, USA)/PBS for 10 min at RT. After PBS rinsing, non-specific binding was blocked with 2.5% normal horse serum (Vector Laboratories, Burlingame, CA, USA) for 1 h at RT. Then, samples were incubated overnight at 4 °C with primary antibodies diluted in 0.1% BSA/PBS against ACTA2 (rabbit, ab32575, Abcam, Cambridge, UK), and anti-TNMD (rabbit polyclonal) generated against TNMD C-terminus (237-317 aa) provided by Prof. Denitsa Docheva (produced in co-operation with Metabion International, Planegg, Germany, PAB 201603-00002). Subsequently, samples were washed in PBS, incubated with 0.3% (v/v) hydrogen peroxide (PanReac AppliChem, Barcelona, Spain) for 15 min at RT, and incubated with anti-rabbit AlexaFluor 488 fluorescent secondary antibody (ThermoFisher Scientific, Waltham, MA, USA) for 1 h at RT. Finally, nuclei and F-actin filaments were counterstained with DAPI and phalloidin for 30 min at RT, respectively. Immunolabeled samples were analyzed through confocal laser scanning microscopy CLSM, TCS SP8, Leica, Wetzlar, Germany) and the quantitative analysis of the immunofluorescence images was performed using ImageJ 1.520 (NIH, Bethesda, MD, USA) software.
4
Quantifying AuNP Endocytosis in U87 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After incubation with PEG-AuNPs or cRGDfK-AuNPs (containing 12 μg/ml Au), cells were rinsed and fixed with PBS and 4% paraformaldehyde respectively. In order to demarcate the outline of the cell, cell staining with acridine orange (AO) was processed at room temperature for 7 min. The cells were then observed using a Leica TCS SP8 CLSM. The endocytosed AuNPs was also quantified by ICP-MS method described above.
U87 cells were pre-incubated with free cRGDfK (50 μg/ml) for 30min prior to the addition of cRGDfK-PEG-AuNPs solution (containing 12 μg/ml Au and 50 μg/ml free cRGDfK). After incubation for another 1 h, the cells were then rinsed, trypsinized and lysed using RIPA solution. Au content was measured using ICP-MS method as mentioned above.
U87 cells were pre-incubated with free cRGDfK (50 μg/ml) for 30min prior to the addition of cRGDfK-PEG-AuNPs solution (containing 12 μg/ml Au and 50 μg/ml free cRGDfK). After incubation for another 1 h, the cells were then rinsed, trypsinized and lysed using RIPA solution. Au content was measured using ICP-MS method as mentioned above.
5
Visualizing Oil-Protein Distribution in Emulsions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The distribution of oil droplets in the FFS and films was observed with a TCSSP8 CLSM (Leica, Germany) according to the report of Cao et al. (2021) . FITC and Nile red were used to detect the protein and oil phase, respectively. The FFS (20 mL) was stained with 50 µL of dye solution containing 0.05% FITC and 0.05% Nile red, deposited on concave glass slides. Then, it was covered with coverslips and examined with a 40× objective. The CLSM images were obtained by selectively exciting them at 552 nm for Nile red and 488 nm for FITC. On the contrary, the stained FFS was used to prepare films according to Section 2.3 . The obtained films were fixed to the slide with a coverslip by neutral balsam and observed with CLSM.
6
Microscopic Imaging of Emulsion Films
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CLSM images of emulsion, FFE and SPI-based emulsion films were captured using a TCS-SP8 CLSM (Leica, Germany). The emulsion, FFE, and emulsion films were stained by FITC solution and Nile red solution as previously reported (Zhao, Ren, Shi, & Weng, 2023b (link)). The protein and oil were detected using laser sources excited at 488 and 552 nm. The diameter distribution of the oil was analyzed by particle size analysis software (Nano measurer 1.2).
7
Allicin Disrupts Trichosporon Biofilms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CLSM analysis was used to assess the effects of allicin on the structure and viability of biofilms. T. asahii cells were cultured in RPMI 1640 medium at 35°C for 6 h in 24-well polystyrene plates, resulting in the adhesion stage of biofilms. The plates were then washed with sterile PBS before biofilm cells were treated by allicin for 24 h at 30°C. After that, the biofilms were stained with a LIVE/DEAD BacLight bacterial viability kit (Thermo Fisher Scientific, USA) for 20 min at 35°C. Viable cells stained with SYTO 9 were observed at 485 nm using a TCS-SP8 CLSM (Leica, Germany), while dead and damaged cells stained with propidium iodide (PI) were observed at 535 nm.
8
Mitochondrial Function Analysis by CLSM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CLSM was performed using a Leica TCS-SP8 CLSM. The JC-1 signal was visualized by excitation at 488 nm and emission at 520–540 nm (green fluorescence) and 595–625 nm (red fluorescence). MitoTracker Red CMXRos (Invitrogen) was visualized by excitation at 578 nm and emission at 580–620 nm. Quantification was performed using Leica confocal software.
9
Multifaceted Characterization of Nanomaterials
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
UV-vis absorption spectra were recorded using a UV-3600 spectrophotometer (Shimadzu, Kyoto, Japan). CD spectra were recorded using the Chirascan Circular Dichroism Spectrometer (Applied Photophysics, London, UK). Features and morphologies of nanomaterials were imaged on JEM1200EX TEM (JEOL, Tokyo, Japan). qRT-PCR was performed and analyzed using the CFX connect software (Bio-Rad, USA). Hydrodynamic size and zeta potential of nanomaterials were measured by Zetasizer Nano ZS (Malvern, UK). Fluorescence spectra were monitored by a Cary Eclipse fluorescence spectrophotometer (Agilent Technologies, Palo Alto, CA). Confocal fluorescence micrographs of cells were acquired on a TCS SP8 CLSM (Leica, Germany). CCK-8 assay was measured with a reader (Thermo Fisher Scientific, USA). Concentrations of all RNA and DNA oligonucleotides were measured by a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). WB images were performed on a ChemiDoc XRS+ imaging system (Bio-Rad, USA). PAGE, SDS-PAGE, and RNA-Denaturing Gel Electrophoresis analysis were imaged on a Bio-Rad ChemiDoc XRS (Bio-Rad, USA). The biodistribution of nanoconjugates was imaged on an IVIS Spectrum in vivo imaging system (PerkinElmer, USA). The FCM analysis was performed on a BD Influx flow cytometer (BD, USA).
10
Quantifying Extracellular Products in Phaeocystis marina Biofilms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Leica TCS SP8 CLSM and the LAS X Version (Leica, Germany) software were used for acquisition of CLSM images. The 63 × (1.4 NA) objective was used for channel mode visualization. Nine sets of images (1024 × 1024 pixels, pixel size = 180.38 nm × 180.38 nm, Image size = 184.52 μm × 184.52 μm, z-step = 0.20 μm) were acquired from three random visual fields in three independent tested P. marina BFs. The threshold value was used to calculate the biovolume of the BFs using the Image J software (National Institutes of Health, United States). Biovolumes represented the amount of the individual components of extracellular products of BFs (μm3).
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!