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Accelerating Cryo-ET Through Improved Vitrification and Monitoring RFA

biohub nyc Chan Zuckerberg Biohub Network

At Biohub, we build the technology to help scientists around the world use AI-powered biology to study how cells operate, organize, and work as part of systems to understand why disease happens and how to correct it. With unprecedented scale of compute, AI research and engineering, and state-of-the-art technology for measuring, imaging, and programming biology, Biohub is leading the first large-scale scientific initiative combining frontier AI with frontier biology.

Biohub seeks to support two-year projects that will improve two crucial areas in cryo-electron tomography (cryo-ET): sample freezing (vitrification) and methods for monitoring the success of the freezing process.

Objectives of the RFA

This RFA will advance sample preparation for cryo-ET imaging through 1) development of new sample vitrification techniques and 2) new methods to monitor and analyze the vitrification process.

Recent advances in cryo-ET technologies have the potential to image the internal architecture of the cell at near-atomic resolution. However, vitrification, the process of rapidly freezing materials to preserve native structures in vitreous ice, is far from an optimized process. Vitrification success rates can be low and often require extensive optimization for each sample type. Poor freezing leads to crystalline ice formation, disrupting structural preservation. There remains a need for improved freezing techniques that enhance the efficiency, reliability, and scalability of sample vitrification to support advanced imaging pipelines.

An inability to assess successful vitrification immediately after freezing also hampers throughput. Currently, vitrification preservation success is typically evaluated during transmission electron microscope (TEM) imaging, which occurs at the end of the complex sample preparation pipeline. Such delays not only hinder sample processing and data collection but also lead to the inefficient use of research funds, equipment, and investigator effort. New methods to monitor vitrification are needed to accelerate cryo-ET sample preparation and increase data collection rates.

Examples of potential areas within the scope of this RFA include, but are not limited to:

Development of novel vitrification approaches or improvements to high-pressure freezing for thick biological samples;
Engineering of sample carriers, interfaces, or instrumentation that enhance heat transfer and freezing reliability;
Real-time or post-freezing methods to assess vitrification quality prior to or during focused ion beam (FIB) milling;
Integrated systems that combine vitrification with monitoring and feedback control; and
Cross-disciplinary approaches from fields such as cryogenics, heat transfer, and materials science that enable improved control and understanding of ice formation in biological specimens.

The physics of ice structure and formation are an area of interest beyond the cryogenic electron microscopy field. We therefore aim to attract proposals from experts in adjacent fields such as ice physics, cryogenics, heat transfer engineering, and materials science, where complementary expertise may unlock novel approaches to vitrification and monitoring challenges.

Out of Scope

Examples of areas outside of the scope of this RFA include:

Vitrification of purified proteins and protein complexes in solution for single-particle cryo-EM;
Methods to address single particle averaging preferred orientation;
Downstream cryo-ET imaging, reconstruction, or data analysis methods;
End-point quality assessment at the TEM stage rather than early or real-time monitoring;
Systems for BSL2+ and above sample preparation;
Incremental optimization of existing vitrification systems without substantive innovation; and
Purely theoretical studies without a clear path to experimental validation in cryo-ET workflows.

AI Based Application Success Predictor

1. Exceptional alignment with the Biohub NY Grand Challenge (highest predictor)

This is the most important predictor.

CZ Biohub NY's stated mission is to:

Harness immune cells for disease detection
Engineer immune cells for disease prevention
Develop technologies for immune-cell monitoring
Create tools for early intervention in disease
Advance immune-cell-based therapeutics

Projects that directly contribute to this Grand Challenge are substantially more competitive.

2. Bold, high-risk, high-reward science (very high predictor)

The Biohub explicitly seeks investigators pursuing:

Transformative ideas
Unconventional approaches
Technological breakthroughs
Ambitious scientific questions

The program was created to fund projects that may be too risky or premature for conventional NIH funding mechanisms.

3. Technology development and tool creation (very high predictor)

A defining feature of Biohub funding is its emphasis on creating enabling technologies.

Strong applications often involve:

Synthetic biology
Immune-cell engineering
Advanced imaging
Single-cell technologies
Computational biology
Genomics
Molecular sensing platforms
New research tools that accelerate discovery

Projects focused solely on descriptive biology are generally less aligned than those producing broadly useful technologies.

4. Interdisciplinary collaboration

Biohub programs are intentionally designed to bring together:

Biologists
Clinicians
Engineers
Computer scientists
Physicists
Data scientists

Successful investigators frequently work across disciplines and demonstrate the ability to build collaborative teams.

5. Potential for transformative impact on human disease

Projects should show a credible path toward:

Earlier disease detection
Disease prevention
Novel therapeutic strategies
Monitoring disease progression
Understanding disease mechanisms

Applications addressing major unmet medical needs are particularly attractive.

6. Investigator creativity and scientific vision

The Biohub Investigator model places substantial emphasis on the investigator rather than only the project.

Past descriptions emphasize support for scientists pursuing:

Their most innovative ideas
New scientific directions
Fundamental unanswered questions

Reviewers often look for visionary investigators capable of driving entirely new research areas.

7. Evidence of scientific excellence and productivity

Although the program encourages bold ideas, successful applicants typically demonstrate:

Strong publication records
Significant scientific contributions
Technical expertise
Leadership in their field
Recognition by peers

Past investigator cohorts have included many highly accomplished early- and mid-career scientists.

8. Commitment to open science and collaborative culture

Biohub strongly emphasizes:

Data sharing
Open dissemination of findings
Cross-institutional collaboration
Participation in investigator meetings

Applicants who embrace collaborative science and knowledge sharing align well with the Biohub philosophy.

9. Ability to catalyze future discoveries

Reviewers favor projects that can:

Create new research platforms
Enable future scientific advances
Generate broadly useful datasets
Support future large-scale research efforts

The goal is often not only solving one problem but creating tools that empower entire research communities.

Specialties

Cell Biology

Eligibility Requirements

Applications may be submitted by domestic and foreign nonprofit and for-profit organizations, public and private institutions, such as colleges, universities, hospitals, laboratories, units of state and local government, companies, and eligible agencies of the federal government. As part of the application process, for-profit organizations may need to provide additional information on the charitable purposes of the proposal. Grants are not permitted to individuals, only to organizations.
An organization may submit more than one application.
Each application should designate one Principal Investigator (PI) as the Coordinating Principal Investigator (Coordinating PI). The Coordinating PI will act as the administrative contact between Biohub and all PIs on the grant. The Coordinating PI must submit the application on behalf of all PIs. The Coordinating PI must be affiliated with the institution submitting the application, and grant funds will be awarded to that institution, which will take responsibility for distributing funds to any other institutions. Note that institutions outside the U.S. may not subcontract to U.S. institutions, so please be mindful when selecting the Coordinating PI/institution.
Each application should have a minimum of one PI (Coordinating PI), but may designate up to three total PIs (one Coordinating PI and up to two Co-PIs).
PIs may only serve as the Coordinating PI on one application, but may serve as a Co-PI on applications different from the one they submitted.
Co-PIs may serve as a Co-PI on multiple applications.
PIs/Co-PIs on one application may be employed at the same or at different institutions.
PIs and Co-PIs must hold a PhD, MD, or equivalent degree.
PIs and Co-PIs must each run laboratories in which they control their budget, their space, and their research. Independence in an academic setting is typically demonstrated by a full-time faculty appointment, a tenure-track position, allocated space, a start-up package, and institutional commitment as defined or verified in a letter from a department chair or equivalent. Independence may be defined differently in other organizations. Note: an upload of the letter or proof of independence is not required at the time of application.
Meta employees, including employees of any subsidiary Meta entities, as well as employees of Chan Zuckerberg Initiative, LLC and Biohub are not permitted to apply.
Biohub reserves the sole right to decide if an applicant and applicant organization meet the eligibility requirements.
Biohub reserves the right to request budget changes prior to award.
Biohub welcomes applications from any country, provided the proposed work is compliant with the United States Treasury Department’s Office of Foreign Asset Control (OFAC) sanctions program. Prior to award, all grant applications will be reviewed for compliance with the United States Treasury Department’s Office of Foreign Asset Control (OFAC) sanctions program, the United States Department of Commerce’s export administration regulations, the Foreign Corrupt Practices Act (FCPA), any other applicable U.S. laws and regulations, and any corresponding laws and regulations in the country where the applicant is based. All grant agreements will also require the grantee to comply with these laws and regulations. For additional information, please refer to the U.S. Treasury Department’s resources, the International Trade Administration’s website on US Export Controls, and the Department of Justice’s website on the FCPA.
While applicants from all countries are welcome to apply, because of required ongoing compliance with U.S. sanctions and export controls, an applicant’s funding eligibility may need to be reassessed if the applicable laws and regulations change at any time. As a result, even if an applicant is eligible to receive funding at the time the application is reviewed, the applicant’s status may change later in the process or during the course of the grant term.While applicants from all countries are welcome to apply, because of required ongoing compliance with U.S. sanctions and export controls, an applicant’s funding eligibility may need to be reassessed if the applicable laws and regulations change at any time. As a result, even if an applicant is eligible to receive funding at the time the application is reviewed, the applicant’s status may change later in the process or during the course of the grant term.

We suggest that you consult your home institution to determine eligibility to apply for this grant and your institutional policy on indirect costs. For questions about eligibility for this award or the application process, please contact us in advance of the proposal deadline at sciencegrants@chanzuckerberg.com. Deadline extensions will not be granted.

Sponsor Details

Sponsor Institute/Organizations: biohub nyc Chan Zuckerberg Biohub Network

Sponsor Type: Corporate/Non-Profit

Address: 615 West 131st Street, 6th Floor, New York, NY 10027

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