Fv10i laser confocal microscope

Manufactured by Olympus

Sourced in Japan

The FV10i laser confocal microscope is a compact and versatile imaging system designed for high-resolution, non-invasive observation of samples. It utilizes laser excitation and confocal detection to provide optical sectioning and improved image contrast compared to traditional widefield microscopy. The FV10i is capable of capturing detailed, three-dimensional images of a variety of specimens.

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Lab products found in correlation

Alexa fluor 488 labeled secondary antibody, by Beyotime (1 mentions) Alexa 594, by Thermo Fisher Scientific (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Alexa 594 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Fluorophore conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Metaxpress software package, by Molecular Devices (1 mentions) Imagexpress micro xls widefield high content analysis system, by Molecular Devices (1 mentions) Image it fix perm kit, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Dapi fluoromount g mounting media, by Southern Biotech (1 mentions) Cellsens digital imaging software, by Olympus (1 mentions) Ix81 inverted fluorescence microscope, by Olympus (1 mentions)

6 protocols using fv10i laser confocal microscope

Cellular Uptake of Labeled sEVs

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When LNCaP cells confluenced to 50–60%, LNCaP-AI + F sEVs labeled with the lipophilic phospholipid membrane dye Dil (Beyotime, China) were added and co-cultured for 48–72 h. After removal of the cell culture medium, the cells were gently washed with PBS three times, fixed with 4% paraformaldehyde for 20 min followed by an overnight incubation with E-cadherin primary antibody (Beyotime, China). The next day, cells were washed twice with PBS and incubated with Alexa Fluor 488-labeled secondary antibody (Beyotime, China) at room temperature for 2 h. Cells were washed again and then stained with DAPI for 5 min. The samples were observed on the FV10i laser confocal microscope (Olympus, Japan) with a 60x UIS2 SAPO air objective.

Lei L., Yu L., Fan W, & Hao X. (2022). Prostate cancer small extracellular vesicles participate in androgen-independent transformation of prostate cancer by transferring let-7a-5p. Heliyon, 8(12), e12114.

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Confocal Imaging of Immunofluorescence

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Immunofluorescence stained images were acquired with an Olympus fv10i laser confocal microscope.

Ayanlaja A.A., Ji G., Wang J., Gao Y., Cheng B., Kanwore K., Zhang L., Xiong Y., Kambey P.A, & Gao D. (2020). Doublecortin undergo nucleocytoplasmic transport via the RanGTPase signaling to promote glioma progression. Cell Communication and Signaling : CCS, 18, 24.

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Immunohistochemistry of Drosophila Eye Tissues

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For immunohistochemistry, larval eye imaginal disc and pupal retinae were dissected in PBS, then fixed in 4% paraformaldehyde/PBS for 20 min and 30 min at 25°C, respectively. After washing with PBS containing 0.3% Triton X-100, the samples were blocked with 10% normal goat serum with PBS containing 0.3% Triton X-100 for 20 min and 30 min at 25°C, respectively. After blocking, primary antibodies were added and incubated for 16 h at 4°C. The following antibodies were used; mouse monoclonal anti-lacZ IgG (diluted at 1:500, DSHB), anti-Dlg aintibody (1:500, DSHB), guinea pig anti-Ziz IgG (1:2,000), anti-Delta (Dl) IgG (1:10, DSHB), anti-diphospho ERK (dpERK) IgG (Sigma) (1:100 dilution) and anti-active Caspase3 IgG (1:500, BD). After washing with PBS containing 0.3% Triton X-100, samples were incubated with secondary antibodies labeled with either Alexa 488 or Alexa 594 (1:400, Invitrogen) for 3 h at 25°C. Alexa 594-conjugated phalloidin (200 units/ml, Invitrogen) was used for the detection of F-actin. After washing with PBS containing 0.3% Triton X-100 followed by washing with PBS, samples were mounted in Vectashield (Vector Lab) and inspected with an Olympus FV-10i laser confocal microscope.

Ozasa F., Morishita K., Dang N.A.S., Miyata S., Yoshida H, & Yamaguchi M. (2017). Drosophila DOCK Family Protein Zizimin Involves in Pigment Cell Differentiation in Pupal Retinae. Cell structure and function, 42(2).

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Vitamin A Storage in iPS-HSCs

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Vitamin A (retinol) storage in iPS-HSCs was analyzed in cells passaged on fibronectin-coated plates or fibronectin-coated microscope slides. The cells were cultured in RPMI 1640 supplemented with 2% B27, 5 μM retinol, and 100 μM palmitic acid for 4 days. Harvested cells were stained with BODIPY493/503, and were analyzed using an FV10i confocal laser microscope (Olympus, Tokyo, Japan). For flow cytometric analysis, cells were dissociated using 0.05% trypsin/0.5 mM ethylenediaminetetraacetic acid (EDTA). The autofluorescence of harvested cells was analyzed by nUV laser (375 nm) and 440/50 filter using a FACS Aria2 cell sorter (Becton Dickinson, Franklin Lakes, NJ, USA)^{44 (link)}.

Miyoshi M., Kakinuma S., Kamiya A., Tsunoda T., Tsuchiya J., Sato A., Kaneko S., Nitta S., Kawai-Kitahata F., Murakawa M., Itsui Y., Nakagawa M., Azuma S., Nakauchi H., Asahina Y, & Watanabe M. (2019). LIM homeobox 2 promotes interaction between human iPS-derived hepatic progenitors and iPS-derived hepatic stellate-like cells. Scientific Reports, 9, 2072.

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Intracellular SREBP-1 Quantification Protocol

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Intracellular SREBP-1 staining was performed using Image-iT® Fix Perm Kit (Thermo Fisher Scientific Inc.). Briefly, cells were fixed and permeabilized in Fixation Buffer and Permeabilization Buffer for 15 min, respectively. The cells were blocked and incubated with a primary monoclonal antibody against SREBP-1 (BioVision Inc., 1:100 dilution). After incubation with fluorophore-conjugated secondary antibodies (Thermo Fisher Scientific Inc.), cells were counterstained with DAPI (Invitrogen, Thermo Fisher Scientific Inc). After washing with PBS, cells were examined using an FV10i confocal laser microscope (Olympus) or the ImageXpress^® Micro XLS Widefield High-Content Analysis System (Molecular Devices). Data analysis was performed using the MetaXpress software package (Molecular Devices).

Chen C.C., Hsu L.W., Huang K.T., Goto S., Chen C.L, & Nakano T. (2017). Overexpression of Insig-2 inhibits atypical antipsychotic-induced adipogenic differentiation and lipid biosynthesis in adipose-derived stem cells. Scientific Reports, 7, 10901.

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Immunofluorescence Analysis of Respiratory and Squamous Markers

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Immunofluorescence was performed on paraffin sections after deparaffinization and rehydration. Phosphate-buffered saline Tween (PBST) was applied prior to antigen retrieval in citrate buffer (pH 6.0). Nonspecific binding was blocked with goat serum (10%) for 10 minutes. Primary antibodies were incubated overnight at 4 ^°C. Slides were washed with PBST, and secondary fluorescent antibodies were applied. DAPI Fluoromount-G mounting media (SouthernBiotech, Birmingham, AB) was used.
Images were visualized using an Olympus IX81 inverted fluorescence microscope or an Olympus FV10i Confocal Laser Microscope (Olympus, Tokyo, Japan). The primary antibodies used are summarized in Table I. The secondary antibodies were purchased from Invitrogen (conjugated to Alexa Fluor 488, Alexa Fluor 549, Alexa Fluor 647). Images were collected at 4×, 40×, and 60× and merged with cellSens digital imaging software (Olympus) and Fiji .^{14 (link)}The trachea was used as the positive control for respiratory markers and the negative control for squamous markers. The esophagus was the positive control for squamous markers and negative control for respiratory markers. Stains performed without primary antibodies were also used as negative controls.

Dowdall J.R., Sadow P.M., Hartnick C., Vinarsky V., Mou H., Zhao R., Song P.C., Franco R.A, & Rajagopal J. (2015). Identification of Distinct Layers Within the Stratified Squamous Epithelium of the Adult Human True Vocal Fold. The Laryngoscope, 125(9), E313-E319.

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