Recommandé
Recommandé
Contenu connexe
Dernier
Dernier (20)
En vedette
En vedette (20)
Biocroi web presentation 07 03-2012
- 1. At the cutting edge of cell based assays Gel Base Bioreactor Technology March 2012 CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 2. The problem CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 3. All chemical and biological processes are influenced by environmental elements such as temperature, pH and chemical composition. CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 4. Environmental Concerns Fluctuations Variations in in thermal dissolved CO2 conditions Cell Based Assays Mechanical Alterations in disturbances water media (Vibrations etc) hydration CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 5. Uncontrolled changes in any of these factors can exert unwanted physical, chemical and/or biological effects on the specimen/sample in question leading to poor reproducibility or disruption of a given scientific and/or manufacturing process CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 6. Negative outcomes Increased Cellular Heterogeneity stress in cell populations Disruption & Poor Reproducibility Aberrant Retardation gene of growth expression CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 7. With the emergence of high throughput and multiplexed biological, chemical and materials screening, the use of multi- welled assay plates has become standard for almost all automated experimental and storage applications. CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 8. However many of these experimental procedures are compromised by positional inequalities between wells on outer and inner locations on the micro-plate. These inequalities or “edge effects” become increasingly apparent as micro-plate densities increase. CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 9. To improve reproducibility when conducting chemical and biological experiments, optimal environmental conditions must be maintained at all times. To achieve this, it is often necessary to deploy costly and cumbersome environmental control systems. These systems dramatically reduce the experimental design flexibility of automated screening platforms and often do not solve the problem. CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 10. Our solution CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 11. Thermal Improved Buffering Cell Growth Phase-lock Technology Prevents Better evaporation well to well consistency CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 12. Side view and plan of gel plate showing construction of wells of plate surrounded by gel. Plate Gel Wells CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 13. Loading in Incubator Un-loading out of incubator Heat, CO2 passes into Gel down thermal and concentration gradient CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 14. A 384 well gel buffered bioreactor CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 15. Top view of the 384 bioreactor plate with gel buffering CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 16. Thermal buffering CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 17. 1.1 1 0.9 0.8 0.7 0.6 Cooling rate reduced tx/t0 % 0.5 0.4 Thermal retention ~ 3 X Improved 0.3 SBS format plate 0.2 0.1 0 0 5 10 15 20 25 30 35 Time (mins) Comparison of cooling between normal and Biocroí plates CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 18. Protection against evaporation CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 19. Wells of 96 welled plates were filled with 100ul serum free culture medium and then maintained in a standard drying oven set at 50ºC for 48 hours. (Data expressed as percentage volume remaining after incubation time. (See plate diagram to identify wells used in all experiments). CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 20. Improved cell growth & reproducibility CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 21. Buffered Plates Normal Plates Wells of buffered and normal 96 well plates were seeded at (equally) low density with cells of immortalised cell line. Prior to maintenance in a standard tissue culture incubator (set at 37C, 95% air/ 5% CO2) for 48 hours. Both buffered and normal plates were placed in middle of incubator side by side. CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 22. Non-insulated 96 well Insulated 96 well 14710 (1871) 16805 (1029) 12.7% 6.12% DRASTIC CV= REDUCTION OF EDGE EFFECTS CV= CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 23. The excellent environmental buffering properties of Biocroí plate technologies allows miniaturisation of cell based assays down to nanoliter volumes. CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 24. High Content Image of cells reverse transfected with SiGlo in a gel buffered 192 nano slide. (Volume 100nl) Miniaturised 1000X As compared to 96- well plate assays normally performed at 100ul CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 25. High Content Image of cells incubated in 100nl in a standard 384-well buffered plate CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 26. Reproducibility of spotting 50 nanoliter siRNA in 384 buffered gel plate CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 27. Dried 50 nanoliter siRNA spot overlain with 100 nanoliter lung carcinoma cells CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 28. Advantages CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
- 29. Increase in Proven reduction throughput in edge effects productivity Substantial Improved data decrease in quality reagent & cell usage CONFIDENTIALITY NOTICE Biocroí Ltd. This document is the property of Biocroí Ltd and contains confidential information. This document shall not be reproduced, Unit 3, 80 The Trinity Centre copied, or used for any purposes without the express written permission of an authorized representative of the company. St James’s Hospital, Dublin 8, Ireland
Notes de l'éditeur
- All chemical and biological processes are influenced by environmental elements such as temperature, pH and chemical composition. Uncontrolled changes in any of these factors can exert unwanted physical, chemical and/or biological effects on the specimen/sample in question leading to poor reproducibility or disruption of a given scientific and/or manufacturing process With the emergence of high throughput and multiplexed biological, chemical and materials screening, the use of multi-welled assay plates has become standard for almost all automated experimental and storage applications. However many of these experimental procedures compromised by positional inequalities between wells on outer and inner locations on the micro-plate. These inequalities or “edge effects” become increasingly apparent as micro-plate densities increase. To improve reproducibility when conducting chemical and biological experiments, optimal environmental conditions must be maintained at all times. To achieve this, it is often necessary to deploy costly and cumbersome environmental control systems which dramatically reduce experimental design flexibility automated screening platforms, often these interventions do not solve the problem.
- All chemical and biological processes are influenced by environmental elements such as temperature, pH and chemical composition. Uncontrolled changes in any of these factors can exert unwanted physical, chemical and/or biological effects on the specimen/sample in question leading to poor reproducibility or disruption of a given scientific and/or manufacturing process With the emergence of high throughput and multiplexed biological, chemical and materials screening, the use of multi-welled assay plates has become standard for almost all automated experimental and storage applications. However many of these experimental procedures compromised by positional inequalities between wells on outer and inner locations on the micro-plate. These inequalities or “edge effects” become increasingly apparent as micro-plate densities increase. To improve reproducibility when conducting chemical and biological experiments, optimal environmental conditions must be maintained at all times. To achieve this, it is often necessary to deploy costly and cumbersome environmental control systems which dramatically reduce experimental design flexibility automated screening platforms, often these interventions do not solve the problem.
- All chemical and biological processes are influenced by environmental elements such as temperature, pH and chemical composition. Uncontrolled changes in any of these factors can exert unwanted physical, chemical and/or biological effects on the specimen/sample in question leading to poor reproducibility or disruption of a given scientific and/or manufacturing process With the emergence of high throughput and multiplexed biological, chemical and materials screening, the use of multi-welled assay plates has become standard for almost all automated experimental and storage applications. However many of these experimental procedures compromised by positional inequalities between wells on outer and inner locations on the micro-plate. These inequalities or “edge effects” become increasingly apparent as micro-plate densities increase. To improve reproducibility when conducting chemical and biological experiments, optimal environmental conditions must be maintained at all times. To achieve this, it is often necessary to deploy costly and cumbersome environmental control systems which dramatically reduce experimental design flexibility automated screening platforms, often these interventions do not solve the problem.