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Fsl confocal microscope

Manufactured by Olympus
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The FSL confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It offers highly sensitive detection, superior image quality, and versatile configuration options to meet the diverse needs of researchers and scientists.

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Lab products found in correlation

Axiovert microscope, by Zeiss (4 mentions) Bodipy tmr x se, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Fluoview fv10 asw3, by Olympus (2 mentions) Bodipy c16, by Thermo Fisher Scientific (1 mentions) Tyramide signal amplification kit, by PerkinElmer (1 mentions) Axio imager 2, by Zeiss (1 mentions) Nestin, by Merck Group (1 mentions) Vimentin, by Merck Group (1 mentions) Chi3l1, by Quidel (1 mentions) α tubulin, by Abcam (1 mentions) Phospho stat3, by Abcam (1 mentions) Anxa2, by BD (1 mentions) Flag tag (flag), by Cell Signaling Technology (1 mentions) Stat3, by Santa Cruz Biotechnology (1 mentions) Bodipy c12, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Biowest (1 mentions)

7 protocols using fsl confocal microscope

1

Lipid Starvation Modulates ZBTB18 Localization

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SNB19 cells were grown in 4-well chamber slides either with 10% FBS DMEM or 10% lipid-depleted FBS (Biowest) DMEM for 48 hours. BTSC cells were grown in 4-well chamber slides in complete Neurobasal medium, as previously described. For lipid starvation SNB19 cells were kept for additional 2 hours either in 10% lipid-depleted FBS DMEM or in 10% FBS DMEM. All cells were then fixed with 4% paraformaldehyde in PBS and processed for the staining. FLAG-ZBTB18 (FL or Nte) was labeled with anti-FLAG M2 monoclonal primary antibody (Sigma #F1804). CTBP2 was labeled with anti-CTBP2 mouse monoclonal antibody (BD Biosciences # 612044). Lipid droplets were stained with anti-Perilipin-3 rabbit polyclonal antibody (Abcam #ab47638) (SNB19 cells), or with 0.5μg/ml Bodipy TMR-X SE (Thermo Fisher Scientific, #D6117) in 150mM NaCl for 10 minutes at room temperature (BTSC475). Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich). Pictures were acquired using a FSL confocal microscope (Olympus).
Masilamani A.P., Schulzki R., Yuan S., Haase I.V., Kling E., Dewes F., Andrieux G., Börries M., Schnell O., Heiland D.H., Schilling O., Ferrarese R, & Carro M.S. (2022). Calpain-mediated cleavage generates a ZBTB18 N-terminal product that regulates HIF1A signaling and glioblastoma metabolism. iScience, 25(7), 104625.
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2

Bodipy-C16 Lipid Uptake Assay

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SNB19 cells were seeded in four-well chamber slides and infected as described above. The cells were then incubated with 50 nM Bodipy-C16 (Thermo Fisher Scientific) in PBS for 15 min at room temperature. Subsequently, the samples were fixed with 4% paraformaldehyde in PBS, counterstained with DAPI, and imaged using a FSL confocal microscope (Olympus).
Ferrarese R., Izzo A., Andrieux G., Lagies S., Bartmuss J.P., Masilamani A.P., Wasilenko A., Osti D., Faletti S., Schulzki R., Yuan S., Kling E., Ribecco V., Heiland D.H., Tholen S., Prinz M., Pelicci G., Kammerer B., Boerries M, & Carro M.S. (2022). ZBTB18 inhibits SREBP-dependent lipid synthesis by halting CTBPs and LSD1 activity in glioblastoma. Life Science Alliance, 6(1), e202201400.
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3

Immunoblotting and Immunostaining Protocol

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Antibodies for immunoblotting and immunostaining are listed in Supplementary Table 5. Secondary labeling of NEFM antibody was performed with Tyramide Signal Amplification Kit (PerkinElmer, Waltham, MA, USA) according to the manufacturer’s instructions. Samples were counterstained with DAPI. Pictures were acquired using an Axiovert Microscope (Zeiss, Oberkochen, Germany) and FSL confocal microscope (Olympus, Tokyo, Japan). Axiovision AXIOVS40 V4.8.0.0 (Carl Zeiss, Oberkochen, Germany) or Fluoview FV10-ASW3.1 (Olympus) software was used for image processing and quantifications.
Masilamani A.P., Ferrarese R., Kling E., Thudi N.K., Kim H., Scholtens D.M., Dai F., Hadler M., Unterkircher T., Platania L., Weyerbrock A., Prinz M., Gillespie G.Y., Harsh IV G.R., Bredel M, & Carro M.S. (2017). KLF6 depletion promotes NF-κB signaling in glioblastoma. Oncogene, 36(25), 3562-3575.
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4

Immunohistochemical Analysis of Tissue Samples

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Histology was performed as previously described.60 (link), 54 (link) Sections were incubated with primary antibodies listed in Supplementary Table 5. Nuclei were counterstained with DAPI. Pictures were acquired using a FSL confocal microscope (Olympus) and AxioImager 2 (Zeiss). Fluoview FV10-ASW3.1 (Olympus) and Adobe Photoshop CS5 software (San Jose, CA, USA) were used for image processing and quantifications. Quantification of Ki67 staining was done using IHC profiler in ImageJ.61 (link)
Masilamani A.P., Ferrarese R., Kling E., Thudi N.K., Kim H., Scholtens D.M., Dai F., Hadler M., Unterkircher T., Platania L., Weyerbrock A., Prinz M., Gillespie G.Y., Harsh IV G.R., Bredel M, & Carro M.S. (2017). KLF6 depletion promotes NF-κB signaling in glioblastoma. Oncogene, 36(25), 3562-3575.
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5

Immunoblotting and Immunostaining Analyses

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The following antibodies were used in immunoblotting analyses: ANXA2 (mouse monoclonal, BD Biosciences), CHI3L1 (rabbit polyclonal, Quidel), FLAG (rabbit, Cell Signaling), and α-tubulin (mouse monoclonal, Abcam), STAT3 (rabbit polyclonal, Santa Cruz), phospho-STAT3 (rabbit, Abcam). Immunostaining was performed using antibodies against: ANXA2 (mouse monoclonal, BD Biosciences), CHI3L1 (rabbit polyclonal, Quidel), Ki67 (mouse monoclonal, Leica), Nestin (rabbit, Millipore), MMP2 (rabbit, Abcam), Vimentin (mouse, Sigma). Pictures were acquired using an Axiovert Microscope (Zeiss) or a FSL confocal microscope (Olympus).
Kling T., Ferrarese R., Ó hAilín D., Johansson P., Heiland D.H., Dai F., Vasilikos I., Weyerbrock A., Jörnsten R., Carro M.S, & Nelander S. (2016). Integrative Modeling Reveals Annexin A2-mediated Epigenetic Control of Mesenchymal Glioblastoma. EBioMedicine, 12, 72-85.
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6

Lipid Uptake Imaging in BTSC475 Cells

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BTSC475 cells were seeded in 4-well chamber slides and infected as described above. The cells were then incubated with 50nM Bodipy-C12 (Thermo Fisher Scientific, #D3822) in PBS for 15 minutes at room temperature. Subsequently, the samples were fixed with 4% paraformaldehyde in PBS, counterstained with DAPI and imaged using a FSL confocal microscope (Olympus).
Masilamani A.P., Schulzki R., Yuan S., Haase I.V., Kling E., Dewes F., Andrieux G., Börries M., Schnell O., Heiland D.H., Schilling O., Ferrarese R, & Carro M.S. (2022). Calpain-mediated cleavage generates a ZBTB18 N-terminal product that regulates HIF1A signaling and glioblastoma metabolism. iScience, 25(7), 104625.
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7

Lipid Droplet Dynamics under Lipid Starvation

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SNB19 cells were grown in four-well chamber slides either with 10% FBS/DMEM or with 10% lipid-depleted FBS (Biowest)/DMEM for 48 h. For lipid starvation, cells were kept for an additional 2 h either in 10% lipid-depleted FBS/DMEM or in 10% FBS/DMEM. All cells were then fixed with 4% paraformaldehyde in PBS and processed for the staining. ZBTB18 was labeled with anti-ZBTB18 rabbit polyclonal primary antibody (#12714-1-AP; Proteintech) and anti-rabbit IgG (H+L) Alexa Fluor 647 (goat; Thermo Fisher Scientific) secondary antibody. Lipid droplets were stained with 0.5 μg/ml Bodipy TMR-X SE (Thermo Fisher Scientific) in 150 mM NaCl for 10 min at room temperature. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich). Pictures were acquired using a FSL confocal microscope (Olympus).
Ferrarese R., Izzo A., Andrieux G., Lagies S., Bartmuss J.P., Masilamani A.P., Wasilenko A., Osti D., Faletti S., Schulzki R., Yuan S., Kling E., Ribecco V., Heiland D.H., Tholen S., Prinz M., Pelicci G., Kammerer B., Boerries M, & Carro M.S. (2022). ZBTB18 inhibits SREBP-dependent lipid synthesis by halting CTBPs and LSD1 activity in glioblastoma. Life Science Alliance, 6(1), e202201400.
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