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30 mm culture plate insert

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The 30-mm culture plate insert is a laboratory equipment item used for cell culture applications. It provides a defined area within a culture plate to contain and support cell growth. The insert is designed to fit standard 35-mm culture dishes.

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Lab products found in correlation

Rhod 2 am, by Thermo Fisher Scientific (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)

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Culture plate inserts (8 citations) Culture insert (2 citations)

3 protocols using 30 mm culture plate insert

1

Porcine Alveolar Macrophages and Tracheal Rings for M. hyopneumoniae Infection

Cited 2 times
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M. hyopneumoniae field strain XLW-1 was isolated from diseased pigs in Jiangsu Province, China. Porcine alveolar macrophages (PAMs) obtained from three 4–5-week-old M. hyopneumoniae-negative piglets, and serologically negative for PRRSV and porcine circovirus type 2 (PCV2), were prepared as described previously [14] (link), [15] (link). Prior to infection, PAMs were mixed and confirmed negative for M. hyopneumoniae, PRRSV, PCV2, pseudorabies virus, and classical swine fever virus by PCR and RT-PCR. RPMI 1640 medium and fetal bovine serum (FBS) were obtained from GIBCO (Invitrogen). The isolated cells were grown and maintained in RPMI 1640 medium containing 10% (v/v) FBS at 37°C with 5% CO2.
In order to simulate natural conditions of M. hyopneumoniae infection, the tracheal ring were infected first. Tracheas were collected as previously described [40] (link). Briefly, the tracheas were excised aseptically from pigs and submerged in chilled PBS. Tracheas were washed with PBS, and transverse sections (approximately 0.5 cm thick) were prepared by making an incision between the tracheal rings. Each tracheal ring was placed in a 30-mm culture plate insert (Millipore, Bedford, Mass.) containing 3 ml of complete medium.
Bin L., Luping D., Bing S., Zhengyu Y., Maojun L., Zhixin F., Yanna W., Haiyan W., Guoqing S, & Kongwang H. (2014). Transcription Analysis of the Porcine Alveolar Macrophage Response to Mycoplasma hyopneumoniae. PLoS ONE, 9(8), e101968.
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2

Glia-Specific Calcium Imaging in Brain Slices

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After a recovery period, slices were bulk loaded with the glia-specific calcium indicator Rhod-2 AM (Life Technologies). A stock solution of Rhod-2 AM was reconstituted in 40 μl of DMSO and stored at −20°C until the day of experiments. The Rhod-2 AM stock solution was used within a week to avoid loss of cell loading capacity. The dye loading dish was prepared using a 30-mm culture plate insert (Millipore, Billerica, MA) placed in a 35-mm dish. One ml of aCSF was added inside and outside of the plate insert prior to moving one or two half slices onto the insert. Two to five μl of Rhod-2 AM stock solution were then dropped directly onto the PVN area of each hemi-slice. Slices were incubated in the dark at room temperature for 1 h, during which 100% O2 was continuously piped into the dish. After the incubation, the brain hemi-slices were washed for 30-240 min in dye-free medium to remove any unincorporated dye and to allow de-esterification of the intracellular acetoxymethyl (AM).
Chen C., Jiang Z., Fu X., Yu D., Huang H, & Tasker J.G. (2019). Astrocytes Amplify Neuronal Dendritic Volume Transmission Stimulated by Norepinephrine. Cell reports, 29(13), 4349-4361.e4.
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3

Glia-Specific Calcium Imaging in Brain Slices

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After a recovery period, slices were bulk loaded with the glia-specific calcium indicator Rhod-2 AM (Life Technologies). A stock solution of Rhod-2 AM was reconstituted in 40 μl of DMSO and stored at −20°C until the day of experiments. The Rhod-2 AM stock solution was used within a week to avoid loss of cell loading capacity. The dye loading dish was prepared using a 30-mm culture plate insert (Millipore, Billerica, MA) placed in a 35-mm dish. One ml of aCSF was added inside and outside of the plate insert prior to moving one or two half slices onto the insert. Two to five μl of Rhod-2 AM stock solution were then dropped directly onto the PVN area of each hemi-slice. Slices were incubated in the dark at room temperature for 1 h, during which 100% O2 was continuously piped into the dish. After the incubation, the brain hemi-slices were washed for 30-240 min in dye-free medium to remove any unincorporated dye and to allow de-esterification of the intracellular acetoxymethyl (AM).
Chen C., Jiang Z., Fu X., Yu D., Huang H, & Tasker J.G. (2019). Astrocytes Amplify Neuronal Dendritic Volume Transmission Stimulated by Norepinephrine. Cell reports, 29(13), 4349-4361.e4.
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