Aromatic Additives for Combined Fluorescence-MALDI Sample Imaging

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Solution Overview

Problem

Current workflows for combining MALDI imaging and fluorescence imaging of biological samples require separate sample preparation steps, leading to increased time and potential sample degradation, and lack integration of MALDI matrices for enhancing fluorescence signals.

Innovation Solution

A method involving the deposition of aromatic additives, such as MALDI matrices, onto the sample before fluorescence imaging, which enhances fluorescence intensity and allows for combined MALDI and fluorescence imaging on the same sample.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If separate sample preparation steps are used for MALDI imaging and fluorescence imaging, then each imaging modality can be optimized independently, but sample preparation time increases and sample degradation risk increases

Engineering Contradiction:

Improveimaging qualityVSAvoidsample preparation time

Core Design Contradiction:

ReliabilityVSLoss of time

Solution Approach 1:

The patent combines MALDI matrix deposition and fluorescence staining into a single integrated sample preparation workflow. The aromatic additives serve dual functions: they act as MALDI matrices for mass spectrometry imaging and simultaneously function as fluorophores for fluorescence imaging. This merging of functions eliminates the need for separate preparation steps, reducing both time and sample handling operations while maintaining optimized performance for both imaging modalities.

Inventive Principle:

Principle #5Merging (Combining)

2Reliability

If separate sample preparation steps are used for MALDI imaging and fluorescence imaging, then each modality can be independently optimized, but the number of sample handling operations increases leading to higher degradation risk

Engineering Contradiction:

Improvesample integrityVSAvoidworkflow complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The aromatic additives employed in this invention exhibit multi-functionality, serving simultaneously as MALDI matrices for desorption/ionization and as fluorophores for optical imaging. This universal application of a single reagent class simplifies the overall workflow by eliminating the need for separate matrix deposition and staining steps, thereby reducing sample handling operations and minimizing degradation risk while maintaining independent optimization capabilities for both imaging modalities.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Measurement precision

If traditional fluorescence imaging is performed without aromatic additives, then the imaging process is simpler, but fluorescence signal intensity is insufficient for sensitive detection

Engineering Contradiction:

Improvefluorescence detection sensitivityVSAvoidsample preparation complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The invention enhances fluorescence detection sensitivity by introducing aromatic additives that modify the optical parameters of the sample. These additives exhibit strong fluorescent properties and can be deposited in controlled amounts to achieve optimal signal intensity. The approach changes the chemical composition parameters of the sample preparation, incorporating aromatic compounds with specific fluorescent characteristics to boost detection sensitivity without significantly complicating the overall workflow.

Inventive Principle:

Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach reduces sample preparation time, minimizes degradation, and enables simultaneous high-quality MALDI and fluorescence imaging, improving molecular detection sensitivity and spatial resolution.

Implementation Method 1

the one or more aromatic additives have an absorbance ranging from about 250 nm to about 500 nm

Methodology Applied

Scientific EffectAbsorbance: Absorption (EM radiation)

Implementation Method 2

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging

Methodology Applied

Scientific EffectLaser desorption/ionization: Laser Ablation

Implementation Method 3

the fluorescence is an autofluorescence intrinsic to the specimen

Methodology Applied

Scientific EffectAutofluorescence: Fluorescence

Data Source

PatentUS20250231190A1Enhanced fluorescence signal through the application of aromatic additives onto the microscopy sample for standard fluorescence, fluorescence microscopy and combined fluorescence maldi microscopy/imaging

Publication Date: 2025.07.17 JOHNS HOPKINS UNIVERSITY

AI summary

Methods for enhancing a fluorescence intensity of a sample by adding an aromatic compound thereto. Workflows for a combined fluorescence-MALDI microscopy/imaging instrument also are disclosed comprising combining MALDI imaging and fluorescence imaging of the same sample in one sample preparation step. The presently disclosed workflow reduces the sampling time to one workday and minimizes sample degradation.