Hybridization of the tissue to gene probes for MERFISH was performed according to instructions provided by Vizgen. Briefly, after aspiration of the 70% ethanol, the 10 μm-thick tissue section mounted onto the MERSCOPE slide was washed with Sample Prep Wash Buffer (Vizgen, PN 20300001) and Formamide Wash Buffer (Vizgen, PN 20300002) while placed in the petri dish, and incubated at 37°C for 30 min in a benchtop incubator (INCU-Line, VWR). The Formamide Wash Buffer was then aspirated, and 50 μL of a custom-designed MERSCOPE gene panel mix was delivered onto the tissue section. A 2 × 2 cm piece of Parafilm M (Fisher Scientific) was carefully placed onto the tissue so that the gene panel mix was evenly spread across the tissue and no bubbles were introduced between the tissue and the film. The petri dish was closed with a lid and sealed with Parafilm M, sterilized with 70% ethanol, and placed in a humidified benchtop incubator (INCU-Line, VWR) set at 37°C for 36–48 h.
Parafilm m
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Parafilm M is a malleable, self-sealing, and non-toxic laboratory film used for sealing and wrapping a variety of containers and samples. It is a versatile product designed to create tight seals and prevent leakage or contamination.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using parafilm m
1
MERFISH Tissue Hybridization Protocol
2
Chick Chorioallantoic Membrane Angiogenesis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Animal
studies were performed in accordance with the guidelines and regulations
laid down in the Animals (Scientific Procedures) Act 1986. CAM model
was carried out in accordance with Home Office Approval, UK (Project
license—PPL P3E01C456). Chicken eggs were acquired from Medeggs
(Norfolk, UK). Eggs were stored in a Hatchmaster incubator (Brinsea,
UK) at 37 °C in a 60% humidified atmosphere and 1 h rotation.
To ensure the maintenance of a humidified environment in the egg incubator,
deionized water (DW) was supplemented every 2 days. Implantation was
carried out after 7 days of incubation. To assess embryo viability
and development, eggs were candled. A window of 1 cm2 was
created with a scalpel onto the egg shell exposing the chorioallantoic
membrane. Hydrogels were implanted, and the window was sealed with
a sterile Parafilm strip (Bemis, Parafilm M, Laboratory Wrapping Film,
Fisher Scientific, UK). Eggs were return to the Hatchmaster incubator
for 7 days (37 °C in a 60% humidified atmosphere) without rotation.
Chalkley scoring was used as previously described3 (link) to quantify infiltration of blood vessels through the implanted
scaffolds. Implants and blank controls were observed in situ under a stereo light microscope. A total of five independent counts
obtained from the number of vessels fitting with the Chalkley graticule
projected onto the samples were registered.
studies were performed in accordance with the guidelines and regulations
laid down in the Animals (Scientific Procedures) Act 1986. CAM model
was carried out in accordance with Home Office Approval, UK (Project
license—PPL P3E01C456). Chicken eggs were acquired from Medeggs
(Norfolk, UK). Eggs were stored in a Hatchmaster incubator (Brinsea,
UK) at 37 °C in a 60% humidified atmosphere and 1 h rotation.
To ensure the maintenance of a humidified environment in the egg incubator,
deionized water (DW) was supplemented every 2 days. Implantation was
carried out after 7 days of incubation. To assess embryo viability
and development, eggs were candled. A window of 1 cm2 was
created with a scalpel onto the egg shell exposing the chorioallantoic
membrane. Hydrogels were implanted, and the window was sealed with
a sterile Parafilm strip (Bemis, Parafilm M, Laboratory Wrapping Film,
Fisher Scientific, UK). Eggs were return to the Hatchmaster incubator
for 7 days (37 °C in a 60% humidified atmosphere) without rotation.
Chalkley scoring was used as previously described3 (link) to quantify infiltration of blood vessels through the implanted
scaffolds. Implants and blank controls were observed in situ under a stereo light microscope. A total of five independent counts
obtained from the number of vessels fitting with the Chalkley graticule
projected onto the samples were registered.
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