In order to analyze the co‐localization of siRNA‐loaded GARP iLNPs with the endosomes or lysosomes at different transfection time points, HepG2‐luc cells were plated into 35 mm dishes at a density of 2 × 105 cells per well. After 24 h, the siRNA‐loaded GARP iLNPs at the final siRNA concentration of 50 nM was added to the dishes. Lipo/Cy5‐siRNA NPs served as a control group. Then the cells were stained and imaged according to above‐mentioned protocol at 1, 3, 5, 8, 10, and 12 h after transfection.
Confocal microscope lsm700
The Zeiss LSM700 is a confocal microscope designed for high-resolution imaging. It features a laser-scanning system and advanced optics for capturing detailed, three-dimensional images of samples. The LSM700 is capable of performing fluorescence imaging and can be used for a variety of applications in life science research and materials science.
Lab products found in correlation
11 protocols using confocal microscope lsm700
Subcellular Localization of siRNA-Loaded GARP iLNPs
In order to analyze the co‐localization of siRNA‐loaded GARP iLNPs with the endosomes or lysosomes at different transfection time points, HepG2‐luc cells were plated into 35 mm dishes at a density of 2 × 105 cells per well. After 24 h, the siRNA‐loaded GARP iLNPs at the final siRNA concentration of 50 nM was added to the dishes. Lipo/Cy5‐siRNA NPs served as a control group. Then the cells were stained and imaged according to above‐mentioned protocol at 1, 3, 5, 8, 10, and 12 h after transfection.
Visualizing Intracellular Salmonella in Caco-2 Cells
Subcellular Localization of NPs/siRNA
Membrane Integrity Assay for S. Typhimurium
Immunofluorescence of FFAR4 in Oocytes
Subcellular Localization of Dually Labeled Nanoassemblies
was used with ASO labeled with Cy5.5 at the 5′ end and rHA
labeled with Cy3. C28/I2 cells were seeded overnight at 10,000 cells/well
in an 8-well chamber slide (Sarstedt, cat# 94.6140.802) at 37 °C,
5% CO2. The cells were then treated with the dually labeled
assembly at 100 nM and incubated for further 4, 6, and 24 h at 37
°C, 5% CO2. Cells were washed with DPBS and then fixed
with formalin 10% (Sigma-Aldrich, cat# HT5014) for 20 min. After further
washing, cells were permeabilized with 0.1% Triton X-100 (Cell Biolabs,
INC., cat# 124102) for 20 min, followed by blocking with 5% bovine
serum albumin (BSA, Sigma-Aldrich, cat# A9647) in DPBS for 1 h. Cells
were washed and incubated with primary rabbit polyclonal antibodies
(Abcam), (anti-EEA1, cat# ab2900, for early endosomes, and anti-Lamp1,
cat# ab24170, for lysosomes) in 5% BSA at 4 °C overnight. The
cells were then washed and incubated with goat antirabbit secondary
antibody, Alexa Fluor 488 (Invitrogen, cat# A11034) for 90 min, followed
by washing and mounting the slide with UltraCruz Aqueous Mounting
Medium with DAPI (Santa Cruz Biotechnology, Inc., cat# sc-24941).
The slides were kept at 4 °C in the dark until visualization
on a Zeiss Confocal microscope LSM 700 (Carl Zeiss MicroImaging).
Images were processed with Zeiss Zen Black 2012 edition.
Multimodal Immune Profiling of Tissue
Brain Tissue Preparation and Immunohistochemistry
Immunohistochemistry was performed as previously described (Liu et al., 2013b (link)). Sections were washed with PBS for 5 min three times, and permeabilized and blocked for 1 hr in 10% donkey serum and 0.2% Triton X-100 before being incubated in the primary antibody in 5% serum and 0.2% Triton X-100 at 4°C overnight. Sections were subsequently washed and stained with Alexa Fluor (Life) secondary antibodies and Hoechst in 5% donkey serum for 1 hr before being washed and mounted onto glass slides with Fluoromount-G mounting solution (SouthernBiotech). The primary antibodies used in this study are listed in
Immunofluorescence Localization of Tribbles Proteins
Podocyte Cytoskeleton Dynamics Imaging
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