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Leica 徕卡
Transforming Organoid & Spheroid 
Imaging with the THUNDER Imager
使用THUNDER成像仪进行成像
Cell 细胞
IMC-0006930]
IMarch. 2024] 2024 年 3 月
From Eye to Insight
从眼睛到洞察力
  • Depth of penetration of light
    光的穿透深度
  • Out-of-focus blur 焦外模糊
  • Lack of contrast 对比度不足
  • Lack of sensitivity (low signal-to-noise ratio)
    灵敏度不足（信噪比低）
  • Phototoxicity & other challenges in performing living specimen experiments
    进行活体实验中的光毒性及其他挑战
  • Need to screen a lot of organoids
    需要筛选大量的器官样本
  • Lack of expertise in analysis or processing tools
    缺乏分析或处理工具方面的专业知识
  • Differing protein expression levels from constructs, requiring a detector with high dynamic range
    来自构建物的蛋白表达水平不同，需要具有高动态范围的探测器

THUNDER Computáional Clearing
计算清除

is an opto-digital method that removes out-of-focus blur in real-time > is fully integrated into the system's data stream
一种光数字方法，可以实时消除焦外模糊 > 完全集成到系统的数据流中
automatically takes all relevant optical parameters into account
自动考虑所有相关的光学参数

Our Solution 我们的解决方案

Imaging Organoids/Spheroids
映射器官样体/球体

Time to Result  结果时间
Rapid acquisition of blur-free images, even deep within thick samples
快速获取无模糊图像，即使在厚样本深处
Extended Range of Applications Fast overviews of whole tissue sections
应用范围扩大，快速查看整个组织切片
Higher Relevance 更高相关性
Imaging and analyzing even
对甚至进行成像和分析
challenging tissue sections with an easy workflow
具有挑战性的组织切片，采用简单的工作流程

Imaging organoids with THUNDER
使用THUNDER成像器官样体

Uncleared human brain organoid (4 colors); Image courtesy by Attria Kavyanifar. M.Sc. (supervised by Prof. Dr. Lie, Prof. Dr. Wiener) University Clinic Erlangen, Germany
未清理的人类大脑器官样体（4 种颜色）；图片由 Attria Kavyanifar 提供。M.Sc.（在 Lie 教授和 Wiener 教授指导下）德国埃尔朗根大学医学院

Extend the Range of Applications to Thicker Samples
将应用范围扩展到更厚的样本

Tumor Spheroids Neurospheres
Widefield 宽场
THUNDER Imager THUNDER成像仪

Increase Relevance: Your Entry to 3D Imaging
提高相关性：您的 3D 成像入口

THUNDER Imager Cell is your versatile entry-level imaging system for a wide range of samples
THUNDER成像细胞是您多样化的入门级成像系统，适用于各种样本
Monolayer Cell Culture 单层细胞培养
3D Cell Culture  3D 细胞培养
Organoids  器官样结构
Spheroids  球体

Reduce Time to Result: Upscaling Your Discovery
缩短结果时间：提升您的发现

THUNDER Imager Cell is your imaging system for a wide range of challenging 3D samples like:
THUNDERImager Cell 是您用于各种具有挑战性的 3D 样本的成像系统，例如：
Spheroids such as tumoroids or neurospheres
瘤球或神经球等球体
Brain or intestine organoids
大脑或肠道器官样本
Also ideal for advanced cell culture experiments such as:
也适用于高级细胞培养实验，如：
Live cell explants or vibratome sections 
Advanced applications like TIRF, FRET 
Perform high resolution tissue imaging on thick samples without struggling to find your region of interest. 
The recently released THUNDER Live tool visualizes a computationally cleared image instantly in the live viewer and allows you to optimize ICC parameters by using live image feedback. 
Your benefits with THUNDER Live: 
  • Get computationally cleared images directly in your live preview. 
  • Reduce the time to optimal results with immediate visual feedback. 
  • Intuitively and quickly find the best THUNDER ICC parameters on the fly while still in live view. 
  • Optimize your image in one simple step without changing any settings. 
  • THUNDER Live's blur-free view also makes selecting significant regions of interest in your sample a breeze, even on thick specimens.
    THUNDER Live 的清晰视野使得即使在厚样本上，选择感兴趣的重要区域也变得轻而易举。
Rabbit Tongue | transversal section of papilla foliata | Acridine Orange stain imaged 2 fluorescence channels DAPI (390nm ex) and FITC (474nm ex)
兔舌|乳头状乳头横截面|吖啶橙染色成像 2 个荧光通道 DAPI（390nm 激发）和 FITC（474nm 激发）

Application example: Tumor Spheroid + THUNDER Live
应用示例：肿瘤球体+THUNDER Live

THUNDER Live reduces the time to search for biological processes in deeper regions of your living sample.
THUNDER Live 减少了在活体样本的深层区域搜索生物过程所需的时间。
Focusing through a thick T47D breast cancer tumor spheroid using THUNDER Live Histone 2B GFP Objective: 20X/0.7
使用THUNDER Live Histone 2B GFP 在 厚的 T47D 乳腺癌肿瘤球体中进行对焦。目标：20X/0.7
Sample Courtesy: 样品提供：
Dr. Nariman Ansari, Leica
纳里曼·安萨里博士，莱卡
Kinetix High End scMos 高端 scMos
camera 相机
K8
K5
Leica LEDs 莱卡 LED 灯
& Synapse & 突触
Attribute 属性 K8
Sensitivity 灵敏度 +++ ++++
QE
Read Noise 读出噪声
Frame rate 帧率 40 fps 40 帧每秒 95 fps 95 帧每秒
 动态范围
Dynamic
range
Cooling 冷却 No 否
Pixel size 像素大小
Resolution 分辨率 4.2 mp 4.2 百万像素 4.2 mp 4.2 百万像素
Applications 应用

使用具有非常高动态范围的固定样本进行成像。使用明亮样本进行活细胞成像。
Imaging/Thunder fixed samples
with very high dynamic range .
Live cell imaging with bright
samples.

活细胞成像低光照Thunder高速成像
Live cell imaging low light
Thunder high speed imaging

Introducing the Leica K8 scientific CMOS camera
推出 Leica K8 科学 CMOS 相机

Sensor type 传感器类型 Scientific CMOS 科学型 CMOS
Sensor size (diagonal) 传感器尺寸（对角线） 18.8 mm 18.8 毫米
Sensor Format 传感器格式 (4.2 Megapixel)  （420 万像素）
Pixel dimensions 像素尺寸
Shutter mode 快门模式 Rolling shutter 滚动快门
Data interface 数据接口 USB 3 Gen2 
Mechanical interface  C -mount 
Exposure range 
Frames per second  95 fps (in 11 bit mode) 
Triggering  Yes, MMCX 
Sensor cooling  at ambient 
Peak quantum effciency 
Dark current  pixel/second 
12 Bit 1.0 e- (Median) 1.1 e-
(RMS)
16 bit 1.6 e- (Median) 1.8 e-
(RMS)
Read noise  45000 e- 
Full well capacity 
Dynamic range  bit, 12-bit, 11-bit, 8-bit
Bit depth 
Binning options 
 
Windows with LAS X
application
Operating system  60 W 
Power consumption 
Operating temperature 
Leica Microsystem I THUNDER Imager 3D Assay & Organoid Imaging | MC--0006930 
Each THUNDER Imager Cell can be a high-speed imaging device whose core components are tightly real-time controlled by Synapse. 
Synapse synchronizes the camera exposure with the actual illumination time. For this reason, Synapse helps to minimize light exposure, photodamage and bleaching by enabling high-speed acquisition. 
Each THUNDER Imager Cell can be equipped with a high-end LED light source, which offers a number of advantages for the user. 
0
Image your samples with high-speed, versatile, and gentler LED light sources. 
3D cell culture samples are typically grown in hydrogels and are comparably far away from the bottom of the sample carrier. 
Long distance water immersion objectives are specifically designed for these types of samples and typically provide much better resolution and higher image brightness as compared to air lenses. 
Keep in mind: 
Image brightness is proportional to the power of the NA 
Matching the RI of specimen and immersion drives image quality and system sensitivity 
Reduce Time to Result: High-Speed Positioning with the Quantum Stage 
Reduce Time to Result: High Speed, More Data Points, Better Statistics 
Reduce Time to Result - Application Workflow Example 
Increase data quality 
Accurately segment your images. 
Open up new experimental options for cell inaging 

THUNDER + Aivia: Two-Color Organoid Cluster Nuclei Segmentation 

$ AIVIA 
Nucleus 
50 um 

THUNDER Offline License 

More time on the THUNDER Imager to perform experiments 

11640708 Processing and Analysis Workstation 

Processing and Analysis Workstation includes PC with preinstalled LAS Office and the following LAS X licenses: 
  • 3D Visualization Basic 
  • Stitching 
  • Ratio Imaging 
  • Colocalisation 
  • Dye Finder 
  • FRET 
    11640709 Processing and Analysis Power Workstation 
following LAS X licenses:
    - 3D Visualization Basic
    - Stitching
    - Ratio Imaging
    - Colocalisation
    - Dye Finder
    - FRET
Processing and Analysis Power Workstation includes PC with preinstalled LAS X Office and the 

Options: 

11889226 THUNDER Offline License OPTION 

11889226 THUNDER Offline is an optional license package exclusively for the following two workstations: 
  • Processing and Analysis Workstation 
  • Processing and Analysis Power Workstation 
Images acquired on a THUNDER Imager with 3D DCV station and LAS X 3.8.0 or higher or Mica system with LAS X 6.2 .0 or higher can be THUNDERed with ICC, SVCC, LVCC. 
Images acquired on a THUNDER Imager with 2D station and LAS X 3.8.0 or higher can be THUNDERed with ICC, CC. 
Please note: THUNDER Offline only works for images deriving from a THUNDER Imager or Mica system 
MICA OPTION 
Learn & Results 
ADDITIONAL LAS X OPTIONS 
2D Analysis 
3D Analysis 
AIVIA OPTIONS 
calling to Aivia options in CPQ 
LAS X Office: Learn and Results 
Resu 
1
.

0 .

LAS X Office: Context Navigator 
Promet statinos 
Organoid/ Spheroid Core 
Fluo turret, mot. 6 pos. with LID 
EFW (4 channel) 
Motorized 6-pos. objective turret with 4x, 10x, 20x and 40x 
Motorized Z-focus drive 
Scanning stage 
LAS X Navigator 
K5 sCMOS camera 
LED5 + pod 
Aivia Go (1 year)  

Application Examples 

Case Studies 
Image Galleries 
Image Gallery: THUNDER Imager T.m.g. posen. cloud the vew of thic sesmples when using camera-base 
Webinars 
"See the Hidden" On Demand 

New e-Book on Organoids & Spheroids! 

Delve into case studies including: 
-Examining 'Brains-In-A-Dish' from induced 

pluripotent stem cells (iPSCs) 
  • Observing 3D cell cultures during development 
-Developing heart pacemaker cells from cardiac spheroids 
Learn more about: 
  • Key considerations for imaging organoids and spheroids 
  • Solutions for examining dynamic processes in organoids and spheroids in real-time 
  • Going deeper into 3D with correlative light 
Biopharma and electron microscopy (CLEM) 
Academia 
versions available 

Additional Resources 

Whitepapers 
From Eye to Insight 
  1. Fluo turret, mot. 6 pos. with LID 
    EFW (>4+ channels) 
    Motorized 6-pos. objective turret with 4x, 10x, 20x and 40x 
    Motorized Z-focus drive 
    Quantum stage inc. multiwheel 
    LAS X Navigator 
    High sensitive fluorescence K8 sCMOS camera 
    LED8 + pod 
    Incubation system (stage top) 
    Aivia Go (1 year)
    Aivia Go（1 年）