US2015099274A1PendingUtilityA1
Method and system for use in monitoring biological material
Est. expiryJun 17, 2032(~5.9 yrs left)· nominal 20-yr term from priority
G01N 2021/3185G01N 21/3504C12Q 1/06C12M 23/36C12Q 1/04G01N 33/49G01N 33/02C12M 41/46G01N 33/497C12M 41/34G01N 21/39G01N 33/4977
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Claims
Abstract
An optical system for determining the concentration of a metabolic gas in a container sealed to biological contamination and enclosing a biological material. The optical system has a broadly tunable coherent infrared light source, a detection module, and a control system connected to the light source and detection module and operates the light source, to receive and analyze the data provided by said detection module, and to process the data indicative of the concentration of said metabolic gas in said sealed container.
Claims
exact text as granted — not AI-modified1 . An optical system for determining the concentration of a metabolic gas in a container sealed to biological contamination and enclosing a biological material, comprising:
(i) a broadly tunable coherent infrared light source adapted to emit (a) a first substantially monochromatic infrared light beam with wavelengths overlapping with an absorption peak of said metabolic gas; and (b) a second substantially monochromatic infrared light beam with wavelengths overlapping with a transmission peak of said metabolic gas or being outside the absorption spectrum of said metabolic gas; (ii) a detection module configured tor detecting said first and said second substantially monochromatic infrared light beams following their passage through a region of interest being part of said sealed container or in fluid communication with it, wherein said region of interest is free of said biological material; and further configured for generating data indicative of transmission of said region of interest to said first and said second substantially monochromatic infrared light beams; and (iii) a control system connectable to said light source and said detection module and suitable to operate said light source, to receive and analyze the data provided by said detection module, and to process the data indicative of the concentration of said metabolic gas in said sealed container.
2 . The optical system of claim 1 , wherein said broadly tunable coherent infrared light source has a tunability range of at least 2 cm −1 .
3 . The optical system of claim 1 , wherein said broadly tunable coherent infrared light source is adapted to emit substantially monochromatic infrared light beam with a spectral width of about 0.7 cm −1 .
4 . The optical system of claim 1 , wherein said broadly tunable coherent infrared light source is adapted to emit substantially monochromatic infrared light beam with a spectral width of about 0.01 cm −1 .
5 . The optical system of claim 1 , wherein said broadly tunable coherent infrared light source is a quantum cascade laser (QCL).
6 . The optical system of claim 1 , wherein said detection module comprises an infrared detector and a lock-in amplifier for improving the signal to noise ratio of said optical system.
7 . The optical system of claim 1 , characterized by a detection sensitivity of said metabolic gas of about 1-10 ppm and a dynamic range of 0-100% relative gas concentration.
8 . The optical system of claim 1 , which is adapted to detect isotopologues of said metabolic gas.
9 . A method for in-situ real-time non-invasive estimation of the level of living cells proliferation and/or growth in a biological material present in a container sealed to biological contamination, said method comprising measuring the concentration of at least one metabolic gas emitted by said living cells according to the following steps:
(i) providing a container sealed to biological contamination and enclosing a biological material; (ii) providing an optical system according to claim 1 ; (iii) measuring the concentration of said at least one metabolic gas emitted by said living cells, which is present in a region of interest, being part of said container or in fluid communication with it, wherein said region of interest is free of said biological material, by:
(a) positioning said region of interest between the broadly tunable coherent infrared light source and the detection module of said optical system;
(b) applying a first substantially monochromatic infrared light beam with wavelengths overlapping with an absorption peak of said at least one metabolic gas and measuring the signal with said detection module;
(c) applying a second substantially monochromatic infrared light beam with wavelengths, overlapping with a transmission peak of said at least one metabolic gas or being outside the absorption spectrum of said at least one metabolic gas and measuring the signal with said detection module;
(d) measuring the concentration of said at least one metabolic gas by processing the results obtained in (b) and (c); and (e) optionally repeating steps (b) to (d) at least one more time;
wherein the concentration of said at least one metabolic gas is an indication of the level of living cells proliferation and/or growth in said sealed container.
10 . The method of claim 9 , wherein the concentration c of said metabolic gas is determined in step (ii)(d) from n measured values of the signal Si (i=1, 2, . . . , n) at different wavelengths λi by utilizing nonlinear minimization of a model S(x, λi) as provided by function s(x) below:
S
(
x
)
=
∑
i
=
1
n
-
1
[
log
(
S
(
x
,
λ
i
)
+
ε
)
S
(
x
,
λ
n
)
)
-
log
(
S
i
+
ε
S
n
)
]
2
where ε is a noise level at the detector, and S(x, λi) is provided by the following equation:
S ( x, λi )= b ∫ λ min λ max f (λ−λ i ) e −α (x+ 0 t 0 ) dλ
where b is a constant f(λ−λ 1 ) is the laser spectral distribution function around the central wavelength λ 1 , αλ is the absorption coefficient, x=ci wherein c is the gas concentration inside the container, l is the pathlength inside the container, c 0 is the concentration of the probed gas outside the container and l 0 is die pathlength outside the container between the infrared source and the detector.
11 . The method of chum 9 , wherein said biological material present in a container sealed to biological contamination is selected from the group consisting of blood components, cell cultures, and microorganisms in a fermentation process.
12 . The method of claim 9 , wherein the distance between the central wavelengths of said first substantially monochromatic infrared light beam and said second substantially monochromatic infrared light beam is below the distance between, two absorption peaks of said at least one metabolic gas.
13 . The method of claim 9 , wherein the spectral width of said first substantially monochromatic infrared light beam is wider than that of the absorption peak of said at least one metabolic gas but narrower than the distance between two absorption peaks of said at least one metabolic gas.
14 . The method of claim 9 , wherein the concentration of said metabolic gas is measured with a sensitivity of about 1-10 ppm and a dynamic range of 0-100% relative gas concentration.
15 . The method of claim 9 , wherein said at least one metabolic gas is selected from the group consisting of carbon dioxide, oxygen, ammonia, hydrogen sulfide, methane, ethane, butane, ethylene, sulfur dioxide, carbonyl sulfide and nitric oxide and isotopologues thereof.
16 . The method of claim 9 , wherein said sealed container to biological contamination is permeable to said at least one metabolic gas and wherein the emission rare of said at least one metabolic gas is determined by applying the following formula:
W ( t )=(ρ( t )−ρ 0 ) vA
where W(t) is the metabolic gas emission rate of enclosed biological material in units kg/s, ρ(ε) is the mass concentration of the metabolic gas in units kg/m 3 at time t, determined in step (iii)(d), ρ 0 is the ambient mass concentration of the gas, v is the membrane permeability coefficient to metabolic gas in units m/s and A is the membrane surface area.
17 . The method of claim 9 , wherein said sealed container to biological contamination is not permeable to said at least one metabolic gas and wherein the concentration of said at least one metabolic gas is determined in step (iii)(d) by applying the following formula:
W ( t )=(ρ( t )=ρ( t− τ)) V/t
where W(t) is the metabolic gas emission rate of enclosed biological material averaged over time interval τ and V is the volume of the container and ρ(t) is the mass concentration of the metabolic gas at time t, determined in step (iii)(d).
18 . A method for detecting a microorganism, contamination in a storage container tor platelets sealed to biological contamination, comprising applying a method according to claim 9 for measuring the concentration of carbon dioxide emitted by said microorganism in said sealed storage container.
19 . A method for monitoring a fermentation process in a fermenter enclosing microorganisms and sealed to biological contamination, comprising monitoring the amount of said microorganisms by applying a method according to claim 9 for measuring the concentration of carbon dioxide emitted by said microorganisms in said sealed fermenter.
20 . A method for monitoring the concentration of living cells in a bioreactor sealed to biological contamination, comprising applying a method according to claim 9 for measuring the concentration of carbon dioxide emitted by said living cells in said bioreactor and optionally correlating said concentration of carbon dioxide to as amount of biomass of said living cells via a linear or robust regression mathematical modelCited by (0)
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