Quantifying the growth of<i>chlamydia suis</i>in cell culture using high‐content microscopy

Puysseleyr, Leentje De; Puysseleyr, Kristien De; Vanrompay, Daisy; Vos, Winnok H. De

doi:10.1002/jemt.22799

Cited by 10 publications

(8 citation statements)

References 32 publications

(31 reference statements)

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“…Fiji Is Just Image J (Fiji) software was used to analyze the images that were obtained with confocal microscopy. Analysis of immunofluorescently labelled cells was done using an updated version of a high content cell analysis script that has been developed earlier [ 71 , 72 ] (CellBlocks.ijm), and which is available upon request. In brief, nuclei were detected in maximum projections of confocal Z-stacks of the DAPI channel after local contrast enhancement to cover for spatial illumination heterogeneity.…”

Section: Methodsmentioning

confidence: 99%

Transient nuclear lamin A/C accretion aids in recovery from vapor nanobubble-induced permeabilisation of the plasma membrane

Houthaeve

Barriga

Stremersch

et al. 2022

Cell. Mol. Life Sci.

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Vapor nanobubble (VNB) photoporation is a physical method for intracellular delivery that has gained significant interest in the past decade. It has successfully been used to introduce molecular cargo of diverse nature into different cell types with high throughput and minimal cytotoxicity. For translational purposes, it is important to understand whether and how photoporation affects cell homeostasis. To obtain a comprehensive view on the transcriptional rewiring that takes place after VNB photoporation, we performed a longitudinal shotgun RNA-sequencing experiment. Six hours after photoporation, we found a marked upregulation of LMNA transcripts as well as their protein products, the A-type lamins. At the same time point, we observed a significant increase in several heterochromatin marks, suggesting a global stiffening of the nucleus. These molecular features vanished 24 h after photoporation. Since VNB-induced chromatin condensation was prolonged in LMNA knockout cells, A-type lamins may be required for restoring the nucleus to its original state. Selective depletion of A-type lamins reduced cell viability after VNB photoporation, while pharmacological stimulation of LMNA transcription increased the percentage of successfully transfected cells that survived after photoporation. Therefore, our results suggest that cells respond to VNB photoporation by temporary upregulation of A-type lamins to facilitate their recovery.

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Section: Methodsmentioning

confidence: 99%

Transient nuclear lamin A/C accretion aids in recovery from vapor nanobubble-induced permeabilisation of the plasma membrane

Houthaeve

Barriga

Stremersch

et al. 2022

Cell. Mol. Life Sci.

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“…The effect of transferrins on C. suis S45 [ 51 ] was examined using McCoy cells (mouse fibroblast cells, CRL-1696 American Type Culture Collection), cultured in Eagle’s minimal essential medium (Thermo Fisher Scientific, Paisley, UK), following standard procedures [ 52 ]. McCoy cells were used, as S45 grows significantly better in these cells compared to, for instance, SK6 and Vero cells [ 53 ]. Titration of the S45 stock was performed by the method used by Spearman and Karber [ 54 ], which determined the TCID 50 .…”

Section: Methodsmentioning

confidence: 99%

“…To analyze the multidimensional image data sets, a dedicated macro script for FIJI image analysis freeware (http://fiji.sc, version 1.53c [57]) was used as described before [53,58]. Briefly, the analysis consists of a stepwise segmentation of regions of interest (nuclei and Chlamydia inclusions), followed by a quantification of their size and number.…”

Section: High Content Microscopy and Image Analysismentioning

confidence: 99%

Transferrins Reduce Replication of Chlamydia suis in McCoy Cells

Puysseleyr

Rybarczyk

et al. 2021

Pathogens

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Chlamydia suis (C. suis) resides in the intestines of pigs and tetracycline-resistant strains are emerging worldwide. Intestinal infections are often subclinical. However, the gut is regarded as a C. suis reservoir and clinical infections have been associated with enteritis, conjunctivitis, pneumonia and reproductive failure. C. suis was found in boar semen and venereal transmission occurred. We studied the anti-Chlamydia suis activity of ovotransferrin (ovoTF) and bovine lactoferrin (bLF). Pre-incubation of C. suis with bLF or ovoTF had no significant effect on overall chlamydia replication (mean fluorescence area) in McCoy cells. The addition of ovoTF to the culture medium had no effect on bacterial replication, but the addition of 0.5 or 5 mg/mL of bLF significantly reduced the inclusion size by 17% and 15% respectively. Egg components are used for cryopreservation of boar semen. When inoculating an ovoTF-containing and Chlamydia suis-spiked semen sample in McCoy cells, a significant reduction in inclusion number (by 7%) and overall replication (by 11%) was observed. Thus, we showed that transferrins possess anti-chlamydial activity. Moreover, ovoTF addition to semen extenders might reduce C. suis venereal transmission. Further research is needed to unravel the mechanisms behind the observations and to enhance the effect of transferrins on C. suis.

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“…Cellular analysis is an important technique used in various fields, such as in biological observations, environmental detections, and medical diagnosis, etc. (De Puysseleyr, De Puysseleyr, Vanrompay, & De Vos, ; Qu, Zhu, & Zhang, ; Stephens & Allan, ). As a classical tool widely used for cellular analysis, optical microscopy is able to provide the direct cellular observations.…”

Section: Introductionmentioning

confidence: 99%

Field of view scanning based quantitative interferometric microscopic cytometers for cellular imaging and analysis

Yan

Wang

2018

Microscopy Res & Technique

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Microimaging is of great significance in the biological and medical fields, since it can realize observations acting as important references for cellular research and disease diagnosis. However, traditional microscopy only offers qualitative sample contours; moreover, it is difficult to reach large-amount sample observations limited by the fixed field of view (FoV). To realize massive cellular measurements quantitatively, three designed quantitative interferometric microscopic cytometers based on the FoV scanning are introduced and compared in details in this article. These devices not only retrieve the quantitative sample phase distributions in the extended FoV, but also provide the detailed information of massive cells, such as cellular volume, area, and roundness. Considering their capabilities as quantitative imaging and large-amount sampling, it is believed that these quantitative interferometric microscopic cytometers (QIMCs) can be potentially adopted in high-throughput cell imaging and statistical analysis for both the biological and medical applications.

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Quantifying the growth ofchlamydia suisin cell culture using high‐content microscopy

Cited by 10 publications

References 32 publications

Transient nuclear lamin A/C accretion aids in recovery from vapor nanobubble-induced permeabilisation of the plasma membrane

Transient nuclear lamin A/C accretion aids in recovery from vapor nanobubble-induced permeabilisation of the plasma membrane

Transferrins Reduce Replication of Chlamydia suis in McCoy Cells

Field of view scanning based quantitative interferometric microscopic cytometers for cellular imaging and analysis

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