เครื่องตัดระดับไมโครด้วยเลเซอร์ภายใต้กล้องจุลทรรศน์ ยี่ห้อ ไลก้า รุ่น LMD6 และ LMD 7 

เราเคลื่อนย้ายเลเซอร์ ไม่ใช่ตัวอย่าง และเราใช้ระบบแรงโน้มถ่วงของโลกในการเก็บตัวอย่างที่สนใจ และนั่นคือเหตุผลที่ระบบ LMD ของเราให้การตัดที่สมบูรณ์แบบ ปราศจากการปนเปื้อน และพร้อมสำหรับนำไปวิเคราะห์

Laser Microdissection (LMD, หรืออีกชื่อหนึ่งคือ Laser Capture Microdissection หรือ LCM) ช่วยให้ผู้ใช้งานสามารถแยกเซลล์เดี่ยว หรือพื้นที่บางส่วนของเนื้อเยื่อออกมาจากตัวอย่างได้ ขับเคลื่อนด้วยการออกแบบเลเซอร์ที่เป็นเอกลักษณ์ พร้อมทั้งซอฟท์แวร์ที่มีประสิทธิภาพ Leica LMD จะช่วยให้ผู้ใช้งานสามารถแยกส่วนที่สนใจออกจากพื้นที่ทั้งหมดของเนื้อเยื่อให้กลายเป็นเซลล์เดียว หรือแม้แต่กลายเป็น subcellular structures เช่น โครโมโซมได้

Leica LMD ถูกใช้อย่างแพร่หลายในงานด้าน genomics (DNA), transcriptomics (mRNA, miRNA), proteomics, metabolomics, และ next generation sequencing (NGS). ซึ่งนักวิจัยทางด้านประสาทวิทยา, การวิจัยทางด้านมะเร็ง หรือนิติวิทยาศาสตร์ก็ได้มีการใช้วิธีนี้ ยิ่งไปกว่านั้น Leica LMD เป็นเครื่องมือที่สมบูรณ์แบบสำหรับการทำ live cell culture (LCC) เพื่อการโคลนนิ่ง และการทำ re-cultivation, manipulation หรือการทำ downstream analysis

Same Principle, Two Systems

LMD is typically used in genomics (DNA), transcriptomics (mRNA, miRNA), proteomics, metabolomics, and even next generation sequencing (NGS). Researchers in neurology, cancer research, plant analysis, forensics or climate research rely on this method. Furthermore, LMD is a perfect tool for live cell culture (LCC), for cloning and re-cultivation, manipulation or downstream analysis.

We move the laser, not the sample. And we use gravity for collection. That is why our LMD systems provide you with perfectly cut, contamination-free, analysis-ready dissectates.

Choose your system

The difference between the Leica LMD6 and Leica LMD7 is the laser. The LMD7 provides higher laser power and more laser controls than the LMD6. This makes the Leica LMD6 the tool for standard applications dissecting soft tissues such as brain, liver, or kidney. And it makes the Leica LMD7 ideally suited to dissect any kind of tissue independent of its size or shape.

Two systems for your discoveries Leica LMD6 for standard tissue dissection Leica LMD7 for higher laser power and more flexibility
Two systems for your discoveries Leica LMD6 for standard tissue dissection Leica LMD7 for higher laser power and more flexibility

We Move the Laser, not the Sample

Try writing your name on a piece of paper by moving the paper instead of the pen. Hard to do? That’s why we move the laser, not the sample for laser microdissection.

Only Leica Microsystems uses high-precision optics to steer the laser beam along the desired cut line on the tissue with prisms. That means that the Leica LMD cuts perpendicularly to the tissue and leaves clean cuts of contamination-free isolates.

Precision. Always.

Laser Adjustment for your Sample

The LMD7’s power, aperture, speed, head current, and pulse frequency settings are tunable to the needs of your sample. For example, thick plant sections require high power, while cutting around fragile neurons will require less power. Thin sections can typically use faster cuts than thick sections.

Advanced cutting modes add enhanced precision to how you will dissect your sample. For example you can draw first and cut afterwards (Draw + Cut), or cut in real-time by mouse click or with a PEN screen (Move + Cut). The Draw + Scan mode is made for ablation from glass, whereas the Laser Screw cuts thick specimen in consecutive rounds. The Final Pulse mode enables you to kick demanding dissectates into the collection vessel. 

Gravity for Cleanliness

Your downstream analysis relies on contaminate-free isolates. This is why Leica LMD systems collect dissectates via gravity. Their unique laser-guidance based dissection method preserves the integrity of your isolates – contact-free and contamination-free.

1, 2, 3 – contamination-free sample!

  1. You choose the area of interest
  2. You move the laser along the area to be cut out
  3. The dissectate drops into the Petri dish and is ready for further analysis

The real asset: Gravity always works.

Leica Laser Microdissection Software

Leica Laser Microdissection Software

The Leica LMD software is an intutive application software designed to streamline the process of selecting settings so that you can focus on obtaining high-quality regions of your sample. Select, dissect, and visualize with ease!

The Leica LMD Software connectivity:

Leica LMD 7 Microdissection Software
Leica LMD 7 Microdissection Software

Automation Options

The ADM (Auto Detection Mode) is a software module which helps collect large amounts of similar dissectates. You just have to encircle a region you are interested in and the software will use this as a template for other regions. For example, Proteomics analysis requires a large amount of sample to be captured, and ADM has provided a great benefit to labs working with 100’s-1000’s of dissectates.

Autofocus can be used before each dissection process to keep the focus of hundreds of ROIs distributed over the slide. Moreover, you can automatically inspect and document all collection devices which were used.

The systematic approach: A raster tool separates your Field of View into a given number of regions. With this function, you can systematically dissect your specimen and collect your dissectate into a variety of collection devices such as 96-Well plates.

Before auto-detectionBefore auto-detectionAfter auto-detection

Interface for Artificial Intelligence

Aivia is our AI-based, image visualization, analysis and interpretation platform. With the help of AI-enhanced tools such as the Pixel Classifier, Aivia can be used to automatically define Regions of interest (ROI) which are destined for Laser Microdissection (LMD). ROIs can be detected by Aivia and imported directly into the LMD software for Microdissection.

Besides Aivia other external software can be used for automatic ROI detection. The LMD software simply needs an XML file containing the ROI information.

Aivia- Automated image segmentation
Aivia- Automated image segmentation

High-Throughput Capability

Need more throughput? The LMD systems can be configured with different kinds of stages. The LMT350 ultra scanning stage provides the fastest, most silent and most precise navigation among them.

The LMT350 is capable of holding collector vessels starting from 0.2 ml tube caps, through 8 and 12 cap-strips up to standard 96-well and 386-well plates*. This enables you to do high-throughput experiments.

Because you don’t need any special material but use regular microtiter plates, these plates can be loaded directly into standard PCR machines.

*max.352 wells addressable

Consumables

A) Specimen

Leica Microsystems offers different slide options for laser microdissection.

Read how to choose the adequate consumable for Laser Microdissection.

B) Collection

Since the Leica LMD systems collect dissectates simply by gravity, you can use standard devices for their collection. Just use common molecular biology reaction devices such as 0.2 or 0.5 ml tube caps – which you already have in your lab. Collection devices can be dry or filled with reaction buffer or culture media for the LMD application.

Leica Laser Microdissection Consumables
Leica Laser Microdissection Consumables

Success is Objective

LMD objectives are designed for the task of enabling flexible laser settings for unique applications. Leica’s development and manufacturing of optics has been among of our core competencies from the start in the 19th century, so you can rely on the performance of our dedicated LMD objectives.

The outstanding imaging performance our objectives provide goes without saying.

Leica Laser Microdissection Objectives
Leica Laser Microdissection Objectives

LCC – Live Cell Cutting

If you work with live cells, you are used to working with inverted microscopes. Live cell cutting works with our LMD systems, even though the Leica LMD systems are based on an upright microscope.

Read more about live cell cutting in our Science Lab article on consumables.

HeLa Cells
HeLa Cells

Combination System: LMD System with THUNDER Imaging

The LMD6 and LMD7 systems can be combined with THUNDER. The base stand of the THUNDER Imager 3D Tissue and LMD systems are the same, thus such a combination offers:

Dedicated Laser Microdissection Objectives

Leica Microsystems offers a portfolio of dry objectives dedicated for Leica LMD systems. These special LMD objectives feature the highest possible UV-transmission and outstanding imaging performance – the Leica SmartCut series (5x–150x).

ObjectiveMag.NAWD (mm)BF, POLFLDIC, PHLMDCISOV
HCX PL FLUOTAR**1.25x0.043.7+++++
PLAN**4x0.126.2+++++++
UVI5x0.1211.7+++++++++
HI PLAN6.3x0.1312.8+++++++++
HCX PL FLUOTAR10x0.311.0++++++++
UVI10x0.252.9+++++++
HCX PL FLUOTAR20x0.46.9++++++++
HCX PL FLUOTAR40x0.63.3–1.9++++++
HCX PL FLUOTAR63x0.72.6–1.8++++++
HCX PL FLUOTAR150x0.90.25++++++

Mag. – magnification; NA – numerical aperture; WD – working distance; BF – brightfield; POL – polarized light; FL – fluorescence; DIC – differential interference contrast; PH – phase contrast; LMD – laser microdissection; CI – cap inspection; SOV – specimen overview; + = suitable; – = not suitable; ++ = dedicated for; +++ = most suitable

** Additionally recommended objectives

Technical Specifications

DissectionDissection and Collection Unit Based on Scanning StageDissection and Collection Unit Based on Motorized StageDissection and Collection Unit Based on Scanning Stage LMT350
Specimen CollectionContact- and contamination-free
Dissection modesDraw & Cut
Move & Cut (direct online cutting)
PtoP (point-to-point)
Draw & Scan (dot dissection scan)
LMD screw (cut subsequently in Z-direction)
Shadow Cuts (parallel draw and dissection)
Serial section cuttingYesNoYes
AVC+ basic
AVC+ professional
Yes
Yes
Yes
Not recommended
Yes
Universal holderYesNoNo
Stage repeatability+/- 1 µm+/- 10 µm<=+/- 0.5 µm
Stage max. travel speed20 mm/s200 mm/s
Stage driveBall bearing screwRack/ pinionLinear motor drives
Holders3x standard slides (25 mm x 76 mm)
Optional:
1x big slide (50 mm x 76 mm) or 2x standard slides (25 mm x 76 mm)
Petri dish (50 mm)
18-well slide (stack)
1x standard slide (25 mm x 76 mm)
Optional:
1x big slide (50 mm x 76 mm) or 2x standard slides (25 mm x 76 mm)
Petri dish (50 mm)
4x standard slides (25 mm x 76 mm) Optional (combinations possible): 1x big slide (50 mm x 76 mm) 2x Petri dishes (50 mm) 2x 18-well Ibidi slide stacks
Collectors4x 0.2 ml standard PCR tubes
4x 0.5 ml standard PCR tubes
Petri dish (50 mm)
Optional:
2x 8-well strips building up a 96-well plate
Height adjustable universal collector (for multi-well slides, 8-strips, 8-strip tube caps, …)
48-well collector (6×8-strip tube caps) (2x for a full 96-well plate)
4x 0.2 ml standard PCR tubes
4x 0.5 ml standard PCR tubes
Optional:
1x 8-well strips building up a 96-well plate
4x 0.2 ml standard PCR tubes 4x 0.5 ml standard PCR tubes 2x Petri dishes (50 mm) Optional: 96-PCR-well plate (non-skirted or semi-skirted) 96-LCC-well plate 2x 18-well Ibidi slides 1 x 8-strip, 1 x 12-strip 1.5 ml tube (with or without filling)
Power supplyCTR6/CTR6 LEDDual SP box
LaserLeica LMD7Leica LMD6
TypeDiode pumped, solid stateDiode pumped, solid state
Wavelength349 nm355 nm
Average pulse energy120 µJ70 µJ
Repetition rate10 – 5000 Hz (free adjustable)80 Hz (fix)
Laser aperture controlYes, continuously adjustable
Free intensity control1-100%
UV offset freely adjustable and saved for each objectiveYes
Laser beam movementVia optics
Microscope stand (fully automated DM6 B)
Transmitted light (TL) axisContrast methodsBF, optional PH, DF, POL, DIC (fully automated)
IlluminationLED (PMax = 15 W ) Lamphousing
AutomationAutomated illumination manager
Automated contrast manager
Constant color intensity control (CCIC) for Halogen TL
CondenserCondenser head S28, 0.55 NA Motorized 7x condenser disk
Motorized polarizer
Fluorescence axisFilter cube turretMotorized 5x or 8x
AutomationFluorescence intensity manager (FIM) for brightness adjustment
Circular and rectangular field diaphragms for eyepiece or camera viewing
Internal filter wheel and motorized Excitation Manager
IlluminationLeica EL6000 (120 W metal halide) or
100 W HBO
Cubes
(individual filters available upon request)
Special cubes (size k) for simultaneous fluorescence and laser application
LMD-BGR
LMD-GFP band pass LMD-GFP long pass LMD-Cy3
LMD-DAPI
LMD-Alexa594
LMD-CFP
LMD-GFP/Cy3
LMD-YFP LMD-Cy5
OperationFocusMotorized:
– 5 electronic ratios
– Includes parfocal function
Memory function for two z-positions
Objective turretMotorized 7x M25 thread including dry and immersion modes
Controls6 programmable function buttons
Leica SmartMove
Controls for z (focus) movement and x, y (stage) movement
4 programmable function buttons
Optional Leica STP8000
Controls for z (coarse and fine focus) and x, y (stage) movement
11 programmable function buttons
Touchscreen with information and control panels
StandDisplayWith integrated touchscreen Leica SmartTouch
Interfaces2 x USB 2.0, 2 x I2C
DimensionsWith scanning stage:
649.6 mm height, 512.0 mm width, 596.5 mm depth
System software (Operating System Microsoft® Windows® 7)
Package includes
(selected features)
DissectionAutomated collection devices and positioning of PCR tubes
Fully automated inspection mode
Multi-cutting over entire slide Save and load drawn shapes
User guidanceWorkflow based graphical user interface
Free scaling of drawn shapes
Save user profiles
Overview images in BF and Fluorescence
ControlFull laser control
Control software of microscope stand
Laser and illumination settings are linked to objectives
InterfacesExport shape list data for Microsoft Excel or OpenOffice
Optional software packages Automated vision control (AVC+ basic) for automated cell recognition within field of view only Automated vision control (AVC+ professional) for fully or semi automated cell recognition over freely defined area
Integrated database to transfer relevant data (laser, microscope and camera; database as optional)