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Cpd030 critical point dryer

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The CPD030 critical point dryer is a laboratory instrument used to dry delicate samples without damaging their structure. It works by transitioning the sample from a liquid state to a gaseous state, avoiding the formation of liquid-gas interfaces that can cause structural damage. The core function of the CPD030 is to facilitate this critical point drying process.

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Lab products found in correlation

Xl30 sem, by Thermo Fisher Scientific (3 mentions) Desk 2 sputter coater, by Thermo Fisher Scientific (3 mentions) Annexin 5 alexa fluor 488 conjugate, by Thermo Fisher Scientific (2 mentions) Quanta 600f sem, by Thermo Fisher Scientific (1 mentions) Vacuum sputter coater, by Denton (1 mentions)

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Critical Point Dryer

4 protocols using cpd030 critical point dryer

1

Listeria Infection Assay in HeLa Cells

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HeLa cells were plated at 5×105 cells per well in 6-well tissue culture plates with etched grid coverslips with imprinted numbers (BELLCO Biotechnology) 24 h prior to infection. Cells were infected with wild type Lm expressing RFP (DPL5538) at an MOI of 100 in DMEM. Bacteria were spun onto cells by centrifugation at 1500 rpm for 5 min. After 60 min of invasion at 37°C, cells were washed three times with phosphate buffered saline (PBS) with Calcium and Magnesium (Wisent #311-420-CL) followed by the addition of growth media containing 50 μg/ml Gentamicin (Wisent #400-130-IG). At 6 h post infection, cells were cooled on ice and washed twice with chilled PBS with Calcium and Magnesium. Annexin V Alexa Fluor 488 Conjugate (Invitrogen) was diluted to 1% (v/v) in chilled PBS with Calcium and Magnesium and added onto the coverslips for 10 min on ice. Cells were washed twice with chilled PBS with Calcium and Magnesium and fixed with 2.5% PFA (EM Sciences #15710) for 30 min at 37°C. Coverslips were imaged by fluorescence microscopy in PBS with Calcium and Magnesium. Subsequently, samples were fixed in 2% glutaraldehyde in cacodylate buffer, rinsed in buffer and dehydrated in a graded ethanol series. The samples were critical point dried in a Bal-tec CPD030 critical point dryer, mounted on aluminum stubs, gold coated in a Denton Desk II sputter coater and examined in an FEI XL30 SEM.
Czuczman M.A., Fattouh R., van Rijn J., Canadien V., Osborne S., Muise A.M., Kuchroo V.K., Higgins D.E, & Brumell J.H. (2014). Listeria monocytogenes exploits efferocytosis to promote cell-to-cell spread. Nature, 509(7499), 230-234.
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2

Listeria Infection Assay in HeLa Cells

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HeLa cells were plated at 5×105 cells per well in 6-well tissue culture plates with etched grid coverslips with imprinted numbers (BELLCO Biotechnology) 24 h prior to infection. Cells were infected with wild type Lm expressing RFP (DPL5538) at an MOI of 100 in DMEM. Bacteria were spun onto cells by centrifugation at 1500 rpm for 5 min. After 60 min of invasion at 37°C, cells were washed three times with phosphate buffered saline (PBS) with Calcium and Magnesium (Wisent #311-420-CL) followed by the addition of growth media containing 50 μg/ml Gentamicin (Wisent #400-130-IG). At 6 h post infection, cells were cooled on ice and washed twice with chilled PBS with Calcium and Magnesium. Annexin V Alexa Fluor 488 Conjugate (Invitrogen) was diluted to 1% (v/v) in chilled PBS with Calcium and Magnesium and added onto the coverslips for 10 min on ice. Cells were washed twice with chilled PBS with Calcium and Magnesium and fixed with 2.5% PFA (EM Sciences #15710) for 30 min at 37°C. Coverslips were imaged by fluorescence microscopy in PBS with Calcium and Magnesium. Subsequently, samples were fixed in 2% glutaraldehyde in cacodylate buffer, rinsed in buffer and dehydrated in a graded ethanol series. The samples were critical point dried in a Bal-tec CPD030 critical point dryer, mounted on aluminum stubs, gold coated in a Denton Desk II sputter coater and examined in an FEI XL30 SEM.
Czuczman M.A., Fattouh R., van Rijn J., Canadien V., Osborne S., Muise A.M., Kuchroo V.K., Higgins D.E, & Brumell J.H. (2014). Listeria monocytogenes exploits efferocytosis to promote cell-to-cell spread. Nature, 509(7499), 230-234.
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3

Zymosan Particle Adhesion Assay

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Twelve millimeter coverslips were etched in piranha solution (3:1 mixture of sulfuric acid and 30% hydrogen peroxide) at room temperature for 15 min, cleaned with deionized (DI) water, and then incubated with 0.01% poly-L-lysine overnight. After rinsing in DI water, the coverslips were placed into a 24-well plate. Zymosan particles were added and incubated on the coverslips for 3 h at room temperature to ensure particle adhesion. Samples were sequentially dehydrated by 30, 50, 75, 90, and 95% ethanol on ice for 5 min each time and 100% ethanol for 5 min, 3 times. Samples were dried using a Leica CPD030 Critical Point Dryer and then sputter-coated with a 10 nm thick film of Pd/Rh alloy using a Denton Vacuum Sputter Coater. Coated samples were imaged using an FEI Quanta 600F SEM with 30 kV voltage, high pressure, and 5–6 mm working distance.
Li W., Wang H., Xu X.G, & Yu Y. (2020). Simultaneous Nanoscale Imaging of Chemical and Architectural Heterogeneity on Yeast Cell Wall Particles. Langmuir : the ACS journal of surfaces and colloids, 36(22), 6169-6177.
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4

Invasion Assay with Salmonella Typhimurium

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Cells were seeded in a 24‐well tissue culture plate at a density of 5 × 104 cells per well and transfected with the indicated siRNA 24 hr later. Invasion was carried out 48 hr after transfection. Cells were infected with S. Typhimurium at the indicated time points. Samples were fixed in 2% glutaraldehyde in cacodylate buffer, rinsed in buffer and dehydrated in a graded ethanol series. The samples were critical point dried in a Bal‐tec CPD030 critical point dryer, mounted on aluminium stubs, gold coated in a Denton Desk II sputter coater and examined in an FEI XL30 SEM.
Boddy K.C., Gao A.D., Truong D., Kim M.S., Froese C.D., Trimble W.S, & Brumell J.H. (2018). Septin‐regulated actin dynamics promote Salmonella invasion of host cells. Cellular Microbiology, 20(10), e12866.
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