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Non-Invasive Optical Sensor Based Approaches for Monitoring Virus Culture to Minimize BSL3 Laboratory Entry.

Ragupathy V, Setty MK, Kostov Y, Ge X, Uplekar S, Hewlett I, Rao G - Sensors (Basel) (2015)

Bottom Line: Routine PPE use involves significant recurring costs.Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs.The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

View Article: PubMed Central - PubMed

Affiliation: LMV/DETTD/OBRR/CBER/FDA, Silver Spring, MD 20993, USA. viswanath.ragupathy@fda.hhs.gov.

ABSTRACT
High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3) laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment (PPE) to observe cell culture conditions. Routine PPE use involves significant recurring costs. Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs. We report here the monitoring of high titer cultures of Human Immunodeficiency Virus-1 (HIV-1) and Herpes Simplex Virus-2 (HSV-2) remotely with the use of optical oxygen sensors aseptically placed inside the cell culture vessel. The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

No MeSH data available.


Related in: MedlinePlus

(A) Fluorometrix Coasters; (B) Fluorometrix Coasters monitoring culture flask; (C) Fluorometrix data recording system.
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sensors-15-14864-f003: (A) Fluorometrix Coasters; (B) Fluorometrix Coasters monitoring culture flask; (C) Fluorometrix data recording system.

Mentions: The sensor (CellPhaseTM, Fluorometrix, currently available from Scientific Industries, Inc. New York, NY, USA) consists of an in dwelling fluorescent oxygen-sensing patch, external optoelectronics and software for user interfaces. A sterile optical patch was placed aseptically in T-75 cm2 flasks (Figure 3A). The patch changes its fluorescence lifetime in response to the changes in oxygen partial pressure. The changes in fluorescence are detected by optoelectronics (Figure 3B). It interrogates the patch with modulated excitation at 470 nm and detects emission at 610 nm. The digitized fluorescent signals are sent to a laptop located outside the BSL3. The fluorescence decay rate is calculated there and converted into an oxygen concentration using manufacturer-supplied calibration codes. The user interface (Figure 3C) displays both the instantaneous value of the dissolved oxygen as well as the time course of the oxygen concentration.


Non-Invasive Optical Sensor Based Approaches for Monitoring Virus Culture to Minimize BSL3 Laboratory Entry.

Ragupathy V, Setty MK, Kostov Y, Ge X, Uplekar S, Hewlett I, Rao G - Sensors (Basel) (2015)

(A) Fluorometrix Coasters; (B) Fluorometrix Coasters monitoring culture flask; (C) Fluorometrix data recording system.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4541811&req=5

sensors-15-14864-f003: (A) Fluorometrix Coasters; (B) Fluorometrix Coasters monitoring culture flask; (C) Fluorometrix data recording system.
Mentions: The sensor (CellPhaseTM, Fluorometrix, currently available from Scientific Industries, Inc. New York, NY, USA) consists of an in dwelling fluorescent oxygen-sensing patch, external optoelectronics and software for user interfaces. A sterile optical patch was placed aseptically in T-75 cm2 flasks (Figure 3A). The patch changes its fluorescence lifetime in response to the changes in oxygen partial pressure. The changes in fluorescence are detected by optoelectronics (Figure 3B). It interrogates the patch with modulated excitation at 470 nm and detects emission at 610 nm. The digitized fluorescent signals are sent to a laptop located outside the BSL3. The fluorescence decay rate is calculated there and converted into an oxygen concentration using manufacturer-supplied calibration codes. The user interface (Figure 3C) displays both the instantaneous value of the dissolved oxygen as well as the time course of the oxygen concentration.

Bottom Line: Routine PPE use involves significant recurring costs.Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs.The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

View Article: PubMed Central - PubMed

Affiliation: LMV/DETTD/OBRR/CBER/FDA, Silver Spring, MD 20993, USA. viswanath.ragupathy@fda.hhs.gov.

ABSTRACT
High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3) laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment (PPE) to observe cell culture conditions. Routine PPE use involves significant recurring costs. Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs. We report here the monitoring of high titer cultures of Human Immunodeficiency Virus-1 (HIV-1) and Herpes Simplex Virus-2 (HSV-2) remotely with the use of optical oxygen sensors aseptically placed inside the cell culture vessel. The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten).

No MeSH data available.


Related in: MedlinePlus