Phase contrast light microscope

Manufactured by Zeiss

Sourced in Germany

The Phase-contrast light microscope is a type of optical microscope that uses phase contrast to enhance the visibility of transparent specimens. It works by converting small differences in refractive index within the sample into differences in brightness, allowing for the observation of cellular structures and other transparent samples without the need for staining.

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Lab products found in correlation

Fresh bone marrow aspirate, by Lonza (2 mentions) Stempro osteogenesis differentiation kit, by Thermo Fisher Scientific (2 mentions) Picogreen assay, by Thermo Fisher Scientific (2 mentions) O cresolphthalein complexone, by Merck Group (2 mentions) Alizarin red s, by Merck Group (2 mentions) Oct compound, by Thermo Fisher Scientific (2 mentions) Crystal violet, by Merck Group (1 mentions)

11 protocols using phase contrast light microscope

Osteogenic Differentiation of hMSCs

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Films for cell culture were sterilized using ethylene oxide for 16 h at 4°C [40 (link)] and stored aseptically until seeding. hMSCs were obtained and cultured as described previously [41 (link)]. hMSCs were isolated from fresh bone marrow aspirates (Lonza, Basel, Switzerland), cultured in Dulbecco’s Modified Eagle Medium (supplemented with 10% fetal bovine serum, 0.1 mM non-essential amino acids, 1 ng/mL bFGF, 1% antibiotic/antimycotic) and seeded at passage 2 [41 (link)]. Cells were seeded at a density of 5,000 cells per cm² in hMSC medium for 3 h, allowing the cells to adhere to the surface. Then, the medium was changed to osteogenic medium StemPro Osteogenesis Differentiation Kit (Gibco, Life Technologies, Grand Island, NY, USA). All cell cultures were performed in an incubator maintained at 37°C and 5% CO₂. Cell growth and shape were monitored using a phase-contrast light microscope (Carl Zeiss, Jena, Germany).

Martín-Moldes Z., López Barreiro D., Buehler M.J, & Kaplan D.L. (2020). Effect of the silica nanoparticle size on the osteoinduction of biomineralized silk-silica nanocomposites. Acta biomaterialia, 120, 203-212.

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Parthenogenetic Activation of Mouse Oocytes

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Vitrified mouse MII oocytes were parthenogenetically activated by culturing in KSOM supplemented with 10 mM SrCl2 for 4 h at 37°C in 5% CO2 in air and then washed three-times with FHM medium. They were cultured in KSOM at 37°C, 5% CO2 in air and monitored under a phase contrast light microscope (Zeiss) for the formation of two-cells, four-cells, morula, and blastocysts at various times for up to 5 days.

Choi J.K., Assal R.E., Ng N., Ginsburg E., Anchan R.M, & Demirci U. (2017). Bio-inspired Solute Enables Preservation of Human Oocytes using Minimum Volume Vitrification. Journal of tissue engineering and regenerative medicine, 12(1), e142-e149.

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hMSCs Osteogenesis Differentiation Protocol

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Human mesenchymal stem cells (hMSCs) were cultured as described previously^{45 (link)}. Briefly, hMSCs were isolated from fresh bone marrow aspirates (Lonza, Basel, Switzerland), cultured in Dulbecco’s Modified Eagle Medium (supplemented with 10% fetal bovine serum, 0.1 mM non-essential amino acids, 1 ng/mL bFGF, 1% antibiotic/antimycotic) and seeded at passage 2. Prior to seeding, hydrogels were extensively extracted with deionized water to remove residual reactants and then incubated in cell culture media for 4 h at 37°C and 5% CO₂. Cells were seeded at a density of 5,000 cells per cm² in hMSC medium for 3 h, allowing the cells to adhere to the surface. Then, the medium was replaced with osteogenic medium StemPro Osteogenesis Differentiation Kit (Gibco, Life Technologies, Grand Island, NY, USA). All cell cultures were performed in an incubator maintained at 37°C and 5% CO₂. Cell growth and shape were monitored using a phase-contrast light microscope (Carl Zeiss, Jena, Germany).

Martín-Moldes Z., Spey Q., Bhatacharya T, & Kaplan D.L. (2022). Silk-elastin-like-protein/graphene-oxide composites for dynamic electronic biomaterials. Macromolecular bioscience, 22(8), e2200122.

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Quantifying Osteogenic Mineralization

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Four and eight weeks post-seeding, films were washed in PBS and osteogenesis was analysed by staining each film with Alizarin Red S (Sigma-Aldrich, St. Louis, MO, USA) to monitor calcium levels in l deposits formed. Cells were fixed with 4% glutaraldehyde for 15min and washed three times with PBS. Next, films were incubated with 2% Alizarin Red S pH 4.2 at room temperature in the dark for 30 min and washed 3 times with PBS. Calcification was monitored using a phase-contrast light microscope (Carl Zeiss, Jena, Germany) equipped with a Sony Exwave HAD 3CCD (Sony Electronics Inc., Park Ridge, NJ, USA) colour video camera. For quantitative analysis of calcium deposition, cells after 4 weeks of culture were first lysed in 0.2% (w/v) Triton X-100 and DNA content was measured using the PicoGreen assay (Molecular Probes, Eugene, OR, USA), according to the protocol of the manufacturer. Samples were measured fluorometrically at an excitation wavelength of 480 nm and an emission wavelength of 528 nm. Calcium was extracted twice with 0.5 mL 5% trichloroacetic acid. Calcium content was determined by a colorimetric assay using o-cresolphthalein complexone (Sigma Aldrich, St. Louis, MO, USA). The calcium complex was measured spectrophotometrically at 575 nm.

Plowright R., Dinjaski N., Zhou S., Belton D.J., Kaplan D.L, & Perry C.C. (2016). Influence of silk-silica fusion protein design on silica condensation in vitro and cellular calcification. RSC advances, 6(26), 21776-21788.

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Oocyte Activation and Fertilization Protocol

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Oocytes were retrieved as previously described²⁵ (link) and cultured in KSOM at 37 °C and 5% CO₂. Half of the retrieved oocytes were treated with hyaluronidase to strip the cumulus granulosa cells, followed by exposure to calcium ionophore A23187 in KSOM media overnight at 37 °C in 5% CO₂. The rest of the oocytes were treated with sperm extracted from the cauda epididymis of C57BL6/J mice. Using a phase-contrast light microscope (Zeiss, Oberkochen, Germany), oocytes were observed for 3 days for any signs of activation or fertilization. Oocytes, granulosa cells, and half of each ovary were snap frozen in liquid nitrogen for subsequent RNA extraction. The other half of the ovary was submerged in optimal cutting temperature (OCT) compound (Thermo Fisher), placed on dry ice to freeze, and stored in −80 °C freezer for future IHC analysis.

Elias K.M., Ng N.W., Dam K.U., Milne A., Disler E.R., Gockley A., Holub N., Seshan M.L., Church G.M., Ginsburg E.S, & Anchan R.M. (2023). Fertility restoration in mice with chemotherapy induced ovarian failure using differentiated iPSCs. eBioMedicine, 94, 104715.

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Morphological Analysis of NRCM

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NRCM morphology was examined under a phase-contrast light microscope (Zeiss Ltd.). Three fields on each well were randomly examined.

Yu W., Kong Q., Jiang S., Li Y., Wang Z., Mao Q., Zhang X., Liu Q., Zhang P., Li Y., Li C., Ding Z, & Liu L. (2024). HSPA12A maintains aerobic glycolytic homeostasis and Histone3 lactylation in cardiomyocytes to attenuate myocardial ischemia/reperfusion injury. JCI Insight, 9(7), e169125.

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Quantifying T. brucei Transporter Impact on T. congolense Growth

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A manual cell count of the cultures was carried out to determine the effect of expression of T. brucei transporters on the growth rate of T. congolense, as described [42 (link)]. Cell lines were counted and adjusted to the same density in 2 mL TCBSF3 medium and incubated in a 24-well plate. The density of each culture was determined by daily manual cell count using a Neubauer cell chamber and a phase-contrast light microscope (Zeiss).

Ungogo M.A., Campagnaro G.D., Alghamdi A.H., Natto M.J, & de Koning H.P. (2022). Differences in Transporters Rather than Drug Targets Are the Principal Determinants of the Different Innate Sensitivities of Trypanosoma congolense and Trypanozoon Subgenus Trypanosomes to Diamidines and Melaminophenyl Arsenicals. International Journal of Molecular Sciences, 23(5), 2844.

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Glioblastoma Cells Colony Formation Assay

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T98, U251, and U87 glioblastoma cells were seeded at a density of 1,000 cells/well into six-well plates and treated with chrysin (0, 30, 60, and 120 µM). The medium and treatment were changed every 3 days. After incubation at 37°C for 10 days, the colonies were washed with PBS, fixed with methanol for 20 minutes, stained with 0.1% crystal violet (Sigma-Aldrich Co) and visualized under a phase-contrast light microscope (Carl Zeiss Meditec AG, Jena, Germany).

Wang J., Wang H., Sun K., Wang X., Pan H., Zhu J., Ji X, & Li X. (2018). Chrysin suppresses proliferation, migration, and invasion in glioblastoma cell lines via mediating the ERK/Nrf2 signaling pathway. Drug Design, Development and Therapy, 12, 721-733.

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Cell Morphology Assessment via Microscopy

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To assess changes in cell morphology, all cell lines (treated and untreated) were examined under phase-contrast light microscope (Zeiss, Berlin, Germany). Furthermore, all cell cultures were monitored for the presence of cytopathic effect (CPE).

Nikolova B., Antov G., Semkova S., Tsoneva I., Christova N., Nacheva L., Kardaleva P., Angelova S., Stoineva I., Ivanova J., Vasileva I, & Kabaivanova L. (2020). Bacterial Natural Disaccharide (Trehalose Tetraester): Molecular Modeling and in Vitro Study of Anticancer Activity on Breast Cancer Cells. Polymers, 12(2), 499.

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Cellular Morphology Under Lipid Treatment

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Cells grown in 24‐well plates were treated with LA at the indicated concentrations for 72 h. Cellular morphology was examined using a phase‐contrast light microscope at magnifications of ×100 and ×400 (Zeiss Ltd., Oberkochen, Germany).

Peng P., Zhang X., Qi T., Cheng H., Kong Q., Liu L., Cao X, & Ding Z. (2020). Alpha‐lipoic acid inhibits lung cancer growth via mTOR‐mediated autophagy inhibition. FEBS Open Bio, 10(4), 607-618.

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