US 2026/0219485 A1Published application

Method for scanning an area with a multi-photon laser scanning microscope and such multi-photon laser scanning microscope

AssigneeFemtonics Kft.

InventorsAndrás FEHÉR, József Balázs RÓZSA, Gergely SZALAY, Katalin ÓCSAI, Gergely KATONA, Máté VERESS, Pál MAÁK, János ORBÁN, Péter PATAI

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Abstract

The invention relates to a method for scanning an area with a multi-photon laser scanning microscope containing acousto-optical deflectors. The method according to the invention comprises scanning the area of a disc lying in an X-Y plane perpendicular to an optical axis of the microscope's objective by applying frequency drive signals to the deflectors, the drive signals having a first term targeting a centre of the disc, and having a second term for scanning a periphery of the disc. The invention further relates to a multi-photon laser scanning microscope containing a drive system configured to perform this method.

Claims17

Dependent claims

Method of scanning an area with a multi-photon laser scanning microscope comprising at least one laser source and a focusing lens system defining an optical Z axis, an optical beam path is provided between the at least one laser source and the focusing lens system, the microscope further comprising first and second acousto-optical deflectors arranged in the optical beam path for deflecting a laser beam exiting the focusing lens system in an X-Z plane defined by the optical Z axis and an X axis perpendicular to the optical Z axis, and third and fourth acousto-optical deflectors arranged in the optical beam path for deflecting the laser beam exiting the focusing lens system in an Y-Z plane defined by the optical Z axis and an Y axis perpendicular to the optical Z axis and the X axis, characterised by scanning a disk lying in a plane perpendicular to an X-Y plane defined by the X axis and the Y axis by guiding the laser beam along the optical beam path from the laser source to the focusing lens system while providing frequency drive signal functions ƒX1(t), ƒX2(t), ƒY1(t), ƒY2(t) to the first, second, third and fourth acousto-optical deflectors defined as

f
X
⁢
1
(
t
)
=
f
0
⁢
X
⁢
1
(
t
)
+
Δ
⁢
f
X
⁢
1
(
t
)
f
X
⁢
2
(
t
)
=
f
0
⁢
X
⁢
2
(
t
)
+
Δ
⁢
f
X
⁢
2
(
t
)
f
Y
⁢
1
(
t
)
=
f
0
⁢
Y
⁢
1
(
t
)
+
Δ
⁢
f
Y
⁢
1
(
t
)
f
Y
⁢
2
(
t
)
=
f
0
⁢
Y
⁢
2
(
t
)
+
Δ
⁢
f
Y
⁢
2
(
t
)
respectively, wherein ƒ0X1(t), ƒ0X2(t), ƒ0Y1(t), ƒ0Y2(t) are frequency drive signal terms for focusing the laser beam in random access point scanning mode to a centre of the disk, the centre having coordinates x, y, z, and ΔƒX1(t), ΔƒX2(t), ΔƒY1(t), ΔƒY2(t) are frequency drive signal terms determining a periphery of the disc and being defined as
Δ
⁢
f
X
⁢
1
(
t
)
=
s
X
⁢
1
·
f
ampX
·
cos
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
Δ
⁢
f
X
⁢
2
(
t
)
=
s
X
⁢
2
·
f
ampX
·
cos
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
Δ
⁢
f
Y
⁢
1
(
t
)
=
s
Y
⁢
1
·
f
ampX
·
sin
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
Δ
⁢
f
Y
⁢
2
(
t
)
=
s
Y
⁢
2
·
f
ampY
·
sin
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
wherein sX1, sX2, sY1, sY2 are selected from values +1 and −1, ƒampX is a first multiplying factor determining a first radius RX of the disc along the X axis, ƒampY is a second multiplying factor determining a second radius RY of the disc along the Y axis, φ is an arbitrary phase shift, and T is a time period required for scanning the disc, and selecting a value of the period T such as to fall between 0.25·Tacc and 1.6·Tacc, wherein Tacc is a filling time of the deflectors.

231 more elements

A multi-photon laser scanning microscope comprising at least one laser source and a focusing lens system defining an optical Z axis, an optical beam path is provided between the at least one laser source and the focusing lens system, the microscope further comprising first and second acousto-optical deflectors arranged in the optical beam path for deflecting a laser beam exiting the focusing lens system in an X-Z plane defined by the optical Z axis and an X axis perpendicular to the optical Z axis, and third and fourth acousto-optical deflectors arranged in the optical beam path for deflecting the laser beam exiting the focusing lens system in an Y-Z plane defined by the optical Z axis and an Y axis perpendicular to the optical Z axis and the X axis, characterised by comprising a drive system configured to scan a disk lying in a plane perpendicular to an X-Y plane defined by the X axis and the Y axis and configured to generate frequency drive signal functions ƒX1(t), ƒX2(t), ƒY1(t), ƒY2(t) for the first, second, third and fourth acousto-optical deflectors defined by

f
X
⁢
1
(
t
)
=
f
0
⁢
X
⁢
1
(
t
)
+
Δ
⁢
f
X
⁢
1
(
t
)
f
X
⁢
2
(
t
)
=
f
0
⁢
X
⁢
2
(
t
)
+
Δ
⁢
f
X
⁢
2
(
t
)
f
Y
⁢
1
(
t
)
=
f
0
⁢
Y
⁢
1
(
t
)
+
Δ
⁢
f
Y
⁢
1
(
t
)
f
Y
⁢
2
(
t
)
=
f
0
⁢
Y
⁢
2
(
t
)
+
Δ
⁢
f
Y
⁢
2
(
t
)
respectively, wherein ƒ0X1(t), ƒ0X2(t), ƒ0Y1(t), ƒ0Y2(t) are frequency drive signal terms for focusing the laser beam in random access point scanning mode to a centre of the disk, the centre having coordinates x, y, z, and ΔƒX1(t), ΔƒX2(t), ΔƒY1(t), ΔƒY2(t) are frequency drive signal terms determining a periphery of the disc and being defined as
Δ
⁢
f
X
⁢
1
(
t
)
=
s
X
⁢
1
·
f
ampX
·
cos
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
Δ
⁢
f
X
⁢
2
(
t
)
=
s
X
⁢
2
·
f
ampX
·
cos
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
Δ
⁢
f
Y
⁢
1
(
t
)
=
s
Y
⁢
1
·
f
ampY
·
sin
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
Δ
⁢
f
Y
⁢
2
(
t
)
=
s
Y
⁢
2
·
f
ampY
·
sin
⁢
(
2
⁢
π
T
⁢
t
+
φ
)
,
wherein sX1, sX2, sY1, sY2 are selected from values +1 and −1, ƒampX is a first multiplying factor determining a first radius RX of the disc along the X axis, ƒampY is a second multiplying factor determining a second radius RY of the disc along the Y axis, φ is an arbitrary phase shift, and T is a time period required for scanning the disc having a value between 0.25·Tacc and 1.6·Tacc, wherein Tacc is a filling time of the deflectors.

231 more elements

Dependencies are inferred from the claim text.

Facts

Application: 19/140,961Filed 18 Dec 2023
Priority: HU P230002622 Dec 2022
Published: 30 Jul 2026
Term ends: Not estimatedThe source records don’t support an estimate.

Classification11 codes

G02B 21/0032
Confocal scanning microscopes or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders: optical…
G01N 21/6458
Spatial resolved fluorescence measurements; Imaging: fluorescence microscopy
G02B 21/0036
Confocal scanning microscopes or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders: scanning…
G02B 21/0076
Optical details of the image generation: arrangements using fluorescence or luminescence