CN109884051A - Harmonic wave confocal micro-measurement method based on image scanning - Google Patents
Harmonic wave confocal micro-measurement method based on image scanning Download PDFInfo
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Abstract
Harmonic wave confocal micro-measurement method based on image scanning belongs to nonlinear optics fields of measurement；In harmonic wave micrometering, replaces conventional confocal to scan the point in micro- with position sensing CCD detection and detect, it can be twice of increase resolution by harmonic wave micrometering.Femto-second laser pulse enters 4f optical system after scanning galvanometer reflects and carries out beam shaping, is then focused at excitation focal beam spot needed for sample interior forms harmonic signal generation by microcobjective.The harmonic signal that sample excitation goes out, again goes into 4f system after being collected by microcobjective.For signal after two separate to dichronic mirror, fundamental frequency signal enters Beam diagnostics camera；Harmonic signal is successively received after confocal pinhole and narrow band filter by position sensing CCD.For each scan position, whole image is recorded by position sensing CCD, and the image of all scan position records is synthesized by algorithm, can promote harmonic wave micro-imaging resolution ratio.
Description
Technical field
The invention belongs to optical microphotograph fields of measurement, relate generally to a kind of for three-dimensional micro- in nano-device and biological sample
The ultraprecise non-contact measurement method of fine texture measurement.
Background technique
It is generated using the nonlinear optical effect of sample itself, such as second harmonic, triple-frequency harmonics generates, and can carry out biology
The micro-imaging of sample unstressed configuration label, the micro-structure detection of nano-device, the diagnosis etc. of pathogenic mechanism.But harmonic wave micro-imaging
The resolution ratio of method is still limited to the optical transfer function and diffraction limit of microscopic system.Many super-resolution fluorescences were aobvious in recent years
Microtechnology, which promotes imaging resolution to diffraction limit, even breaks through diffraction limit.Wherein Structured Illumination is micro- for one of which
Important technology.In Structured Illumination is micro-, sample is illuminated and is inspired by the excitation intensity distribution Jing Guo spatial modulation
Signal be collected by traditional wide field imaging device.By different location and direction be mobile and rotation excitation intensity mode,
And each wide-field image is recorded, the image after synthesis can be nearly twice by increase resolution.But structure light is needed to illuminating bundle
It is modulated and imaging device is relative complex.
Micro imaging method of the another kind based on Structured Illumination thought is image scanning micro imaging method.Image scanning
Micro imaging method, which focuses the laser under diffraction limit, regards a kind of special Structured Illumination as, carries objective lens numerical hole
The permitted all possible spectrum mode of diameter.A detection is changed to by this method on the basis of conventional confocal scans microscopic method
Position sensing CCD detection, for each scan position, whole image is recorded.The image that all records are synthesized by algorithm, can
To promote micro-imaging resolution ratio.Image scanning micro imaging method is integrated as further mentioning with harmonic wave micro-imaging technique
It rises harmonic wave micro-imaging resolution ratio and provides a new technological approaches.
Summary of the invention
The present invention devises a kind of harmonic wave confocal micro-measurement method based on image scanning, with position sensing CCD mostly as
The photomultiplier tube point that element detection replaces conventional harmonic to scan micro- middle use detects, to realize harmonic wave micro-imaging indirectly
The Structured Illumination of technology, while keeping imaging system simpler compared to traditional structure lighting system and easy to operate.It will be every
The entire harmonic signal image that a scan position is detected synthesizes final harmonic image by algorithm, can be humorous by tradition
The micro- imaging resolution of wave is promoted nearly twice.
The object of the present invention is achieved like this:
Harmonic wave confocal micro-measurement method based on image scanning, replaces conventional harmonic confocal with position sensing CCD detection
Scan the point detection in micro-.Femto-second laser pulse enters 4f optical system after scanning galvanometer reflects to carry out through beam shaping,
Then excitation focal beam spot needed for sample interior forms harmonic signal generation is focused at by microcobjective.Sample excitation goes out humorous
Wave signal again goes into 4f system after being collected by microcobjective.For signal after two separate to dichronic mirror, fundamental frequency signal enters light
Beam diagnoses camera；Harmonic signal is successively received after confocal pinhole and narrow band filter by position sensing CCD.It is described based on
The harmonic wave confocal micro-measurement method of image scanning it is characterized in that by with structure light thought image scanning microscopic method with
Harmonic wave micro-imaging technique organically combines.
The above-mentioned harmonic wave confocal micro-measurement method based on image scanning, it is characterised in that with position sensing CCD mostly as
The photomultiplier tube point that element detection replaces conventional harmonic to scan micro- middle use detects.
The above-mentioned harmonic wave confocal micro-measurement method based on image scanning, it is characterised in that position is scanned for each
It sets, whole image is recorded by position sensing CCD, and is synthesized the harmonic image that all scan positions are recorded by algorithm
Final high-resolution harmonic image.
The above-mentioned harmonic wave confocal micro-measurement method based on image scanning, it is characterised in that using confocal pinhole to harmonic wave
Signal carries out apodization.
Due to replacing conventional harmonic with the more pixel detections of position sensing CCD in harmonic wave micro imaging method of the invention
The photomultiplier tube point detection of micro- middle use is scanned, so that the Structured Illumination of harmonic wave micro-imaging technique is realized indirectly,
Keep imaging system simpler compared to traditional structure lighting system simultaneously and easy to operate.Each scan position is detected
Entire harmonic signal image final harmonic image is synthesized by algorithm, imaging resolution that can be micro- by conventional harmonic
It is promoted nearly twice.
Detailed description of the invention
Fig. 1 is the harmonic wave confocal micro-measurement method schematic diagram based on image scanning.
Specific embodiment
Embodiment of the invention is described in detail below in conjunction with attached drawing.
The harmonic wave confocal micro-measurement method schematic diagram based on image scanning of the present embodiment is as shown in Figure 1.Femtosecond laser
Pulse enters 4f optical system after scanning galvanometer reflects and carries out then being focused at sample interior by microcobjective through beam shaping
Form the excitation focal beam spot needed for harmonic signal occurs.The harmonic signal that sample excitation goes out, it is another after being collected by microcobjective
It is secondary to enter 4f system.For signal after two separate to dichronic mirror, fundamental frequency signal enters Beam diagnostics camera；Harmonic signal successively passes through
It is received after confocal pinhole and narrow band filter by position sensing CCD.For each scan position, whole image is quick by position
Feel CCD record, and the harmonic image that all scan positions are recorded is synthesized by algorithm by final high-resolution harmonic wave figure
Picture.
Claims (4)
1. the harmonic wave confocal micro-measurement method based on image scanning, replaces conventional harmonic is confocal to sweep with position sensing CCD detection
Retouch the point detection in micro-.Femto-second laser pulse enters 4f optical system after scanning galvanometer reflects connect through beam shaping
By microcobjective be focused at sample interior formed harmonic signal occur needed for excitation focal beam spot.The harmonic wave that sample excitation goes out
Signal again goes into 4f system after being collected by microcobjective.For signal after two separate to dichronic mirror, fundamental frequency signal enters light beam
Diagnose camera；Harmonic signal is successively received after confocal pinhole and narrow band filter by position sensing CCD.It is characterized in that will
Image scanning microscopic method and harmonic wave micro-imaging technique with structure light thought organically combine.
2. the harmonic wave confocal micro-measurement method according to claim 1 based on image scanning, it is characterised in that use position
Set the photomultiplier tube point detection that the sensitive more pixel detections of CCD replace conventional harmonic to scan micro- middle use.
3. the harmonic wave confocal micro-measurement method according to claim 1 based on image scanning, it is characterised in that for
Each scan position, whole image are recorded by position sensing CCD, and by algorithm all scan positions are recorded it is humorous
Wave image synthesizes final high-resolution harmonic image.
4. the harmonic wave confocal micro-measurement method according to claim 1 based on image scanning, it is characterised in that use
Confocal pinhole carries out apodization to harmonic signal.
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Cited By (3)
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| CN110638424A (en) * | 2019-09-19 | 2020-01-03 | 哈尔滨工业大学 | Scanning light sheet harmonic wave microscopic imaging method and device |
| CN110664369A (en) * | 2019-09-19 | 2020-01-10 | 哈尔滨工业大学 | Self-adaptive confocal line scanning harmonic microscopic imaging method and device |
| CN111879740A (en) * | 2020-07-14 | 2020-11-03 | 哈尔滨工业大学 | All-optical super-resolution microscopic device based on photon reset technology |
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Application publication date: 20190614 |