Two-band optical comparator for use with chopped CW singlet oxygen monitor

• US 4,827,938 A
• Filed: 08/09/1988
• Issued: 05/09/1989
• Est. Priority Date: 05/21/1986
• Status: Expired due to Fees

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Parker, John G.
• Primary Examiner(s): Howell, Kyle L.
• Assistant Examiner(s): Hanley, John C.
• Application Number:
  US07/230,706
• Time in Patent Office: 273 Days
• Field of Search: 128/633-635, 128/654, 128/664, 128/665, 128/395-398, 128/362, 128/303.1
• US Class Current: 600/317
• CPC Class Codes: A61B 2017/00057 Light
• CPC Class Codes: A61N 2005/0644 Handheld applicators
• CPC Class Codes: A61N 5/062 Photodynamic therapy, i.e. excitation of an agent

Abstract

An improved apparatus and method for detecting singlet oxygen emission produced during photoradiation is disclosed. The invention utilizes a two-band optical comparator to detect optical emissions. The optical comparator comprises an in-band filter that passes light in the 1270 nm singlet oxygen emission band and an out-of-band filter that passes light outside of the 1270 nm singlet oxygen emission band. The signal detected when the out-of-band filter is used is composed essentially of a fluorescence emission. This detected signal is used to synchronize an electronic reference signal. The electronic reference signal is used by a signal processing means to isolate a phase delayed singlet oxygen emission component which appears with a fluorescence emission component in the 1270 nm singlet oxygen emission band.

Claims

1. An apparatus for detecting singlet oxygen emission produced during photoradiation, comprising:

an optical excitation means for illuminating a particular portion of a medium containing a photodynamic sensitizer with chopped optical radiation of a wavelength capable of at least causing generation of emission around 1270 nm, wherein said emission includes a first emission component and a second emission component out of phase with said first emission component;

an means for collecting emission from said medium and for sequentially and selectively transmitting a first band of emissions containing a first spectral portion of said emission adjacent to but not including the wavelength of 1270 NM, and then a second band containing a second spectral portion of said emission including said 1270 nm wavelength;

an electro-optical means for detecting emission collected and filtered by said for collecting means, detection of emissions transmitted in said first spectral band producing a first output electrical signal responsive essentially to said the intensity of first portion of said emission, detection of emissions transmitted in said second spectral band producing a second output electrical signal responsive to the intensity of;

said second portion of said emissions;

a means for generating an electrical reference signal of the same frequency as said chopped optical excitation radiation,

a means for adjusting the phase of said electrical reference signal until it is in phase with said first output electrical signal detected in said first band; and

,

a signal processing means for separating said second emission component from said second output electrical signal by determining the component of said second output electrical signal in said second band that is about 90°

out of phase with said synchronised electrical reference signal, whereby said component of said second output signal is a measure of singlet oxygen emission.

2. The apparatus of claim 1, wherein said means for collecting comprises:

a first filtering means for blocking light outside said first band of emissions;

a second filtering means for blocking light outside said second band of emissions, and,

a means for selectively and sequentially placing said first filtering means and said second filtering means so as to alternatively transmit said first band of emissions or said second band of emissions.

3. The apparatus of claim 1, wherein said electrooptical means includes a preamplifier means for amplifying said detected emissions.

4. The apparatus of claim 1, wherein said signal processing means is a lock-in amplifier receiving as inputs said phase adjusted electrical reference signal and said first or second output electrical signals.

5. A signal processing method for detecting singlet oxygen emission produced during photoradiation, said method comprising the steps of:

illuminating a portion of a medium containing a photodynamic sensitizer with chopped optical radiation having a wavelength capable of at least causing generation of singlet oxygen emission;

collecting luminescence emitted from said medium;

filtering said collected luminescence with a first filter to selectively pass wavelengths in a first spectral band, said first band being spectrally adjacent but not including a wavelength of 1270 nm;

generating an electrical reference signal of the same frequency as said chopped optical radiation;

detecting said emission in said first spectral band and producing a first output electrical signal;

comparing the phase of said first output electrical signal detected in said first band with said electrical reference signal;

adjusting the phase of said reference signal to make it synchronous with said first output electrical signal;

removing said first filter and replacing it with a second filter to selectively pass wavelengths of luminescence in a second spectral band that includes the wavelength of 1270 nm but excludes wavelengths of luminescence outside of the 1270 nm band;

detecting said wavelengths of luminescenc in said second band and producing a composite output signal including a fluorescence emission signal component and a singlet oxygen emission signal component out of phase from said fluorescence emission signal component; and

,

processing said composite output signal to separate the singlet oxygen emission signal component from said fluorescence emission signal component based on their relative phase difference, by determining the component of said composite output signal that has a predetermined phase difference from the synchronized reference signal.

6. The method of claim 5, wherein said step of adjusting the phase of said reference signal comprises the steps of:

shifting the phase of said electrical reference signal by 90°

, thereby producing a quadrature of said electrical reference signal;

inputting both said quadrature of said electrical reference signal and said first output electrical signal detected in said first band into a synchronous detector; and

,

adjusting the phase of said quadrature of said electrical reference signal until the output from said synchronous detector is zero.

7. The method of claim 5, wherein said step of determining involves the step of:

separating a component of said composite electrical signal detected in said second band that is about 90°

out of phase from said synchronized electrical reference signal, said separated component representing the singlet oxygen emission signal component.

8. The method of claim 7, wherein said separating step further involves the steps of:

shifting the phase of said synchronized electrical reference signal by 90°

thereby generating a quadrature electrical reference signal; and

,

inputting said quadrature electrical reference signal and said composite electrical signal detected in said second band to a synchronous detector, the output of said synchronize detector representing the singlet oxygen emission signal.